Several MPN models for serial dilutions with suppressed growth at low dilutions

Most probable number (MPN) estimates of microbial concentrations from serial dilutions employ a usual model of probabilities of the various possible outcomes. Outcomes challenge the usual model when they occur at markedly different rates from what the usual model predicts. Too many outcomes that suggest suppression of growth at low dilutions have occurred in some foods. These challenge the usual model and suggest new models. This paper provides four new models: growth suppression by a toxicant released from the food product by sample preparation; interference by a non-target species of microbe; disaggregation of clumped microbes at a single dilution step; disaggregation spread evenly over several dilution steps. The assumptions and discussion of how to find the maximum likelihood estimate for the parameters are given.     

Effect of some hydrocolloids on the rheological properties of different formulated ketchups

Five different hydrocolloids (tragacanth gum, guar gum, carboxy methyl cellulose, xanthan gum and locust bean gum) were added, at levels of 0, 0.5, and 1 g/100 g (w/w), respectively, to three different formulated ketchups which were processed from cold-break tomato paste dilutions, having total soluble solid (TSS) contents of 7.5, 10, and 12.5 g/100 g (w/w), in sequence, and the effect of these hydrocolloids on the rheological properties of tomato ketchups was investigated using a viscometer with smooth surface wide-gap coaxial cylinders. All hydrocolloids increased the consistency of the tested samples; however, guar gum and locust bean gum caused the maximum increase, followed by xanthan gum, tragacanth gum and carboxy methyl cellulose (CMC). Both the ketchup formulation and the hydrocolloid concentration were found to affect the consistency of ketchups. The highest consistency index was obtained by processing dilutions with a TSS content of 12.5%, and the addition of hydrocolloids at the level of 1%. The fluidity of the ketchups decreased with both the addition of all hydrocolloids and the increase in hydrocolloids concentration. Furthermore, the fluidity of the ketchups was also affected by ketchup formulation, and it was found to be the lowest for the samples prepared from the tomato paste dilutions having a TSS content of 12.5%.     

The practical analysis of food: The development of Sakalar quantification table of DNA (SQT-DNA)

Practical and highly sensitive Sakalar quantification table of DNA (SQT-DNA) has been developed for the detection% of species-specific DNA amount in food products. Cycle threshold (Ct) data were obtained from multiple curves of real-time qPCR. The statistical analysis was done to estimate the concentration of standard dilutions. Amplicon concentrations versus each Ct value were assessed by the predictions of targets at known concentrations. SQT-DNA was prepared by using the percentage versus each Ct values. The applicability of SQT-DNA to commercial foods was proved by using sausages containing varying ratios of beef, chicken, and soybean. The results showed that SQT-DNA can be used to directly quantify food DNA by a single PCR without the need to construct a standart curve in parallel with the samples every time the experiment is performed, and also quantification by SQT-DNA is as reliable as standard curve quantification for a wide range of DNA concentrations.     

Water quality and taste sensitivity for basic tastes and metallic sensation

The aim of the study was to investigate the influence of water quality on taste sensitivity for basic tastes and metallic sensation. To familiarize subjects with sensory analysis, they were asked to recognize tastes, distinguish between different concentrations of basic tastes, and realize their own taste sensitivity. The International Standard ISO 3972 describes a set of tests to measure taste sensitivity. A “neutral, tasteless, still and odourless water, preferably of known hardness” is recommended for preparing the dilutions and for rinsing the mouth. The standard also recommends a series of dilutions for acid (citric acid), bitter (caffeine), salty (sodium chloride), sweet (sucrose), umami (monosodium glutamate) and metallic (iron(II)sulfate). The question arises if water quality affects taste sensitivity.MethodsTwenty-three female subjects (mean age: 22), who were recruited at the University of Applied Sciences Hamburg, Faculty of Life Sciences, were tested. Deionized water, spring water and tap water were used for taste dilutions. In each experiment, subjects were initially familiarized with the basic tastes.ResultsIn comparison to taste thresholds in spring water, the recognition thresholds are higher using tap water, and lower using deionized water. When using deionized water, sour was detected by all subjects at the lowest concentrations. Iron(II)sulfate could best be matched to metallic sensation in deionized water. Results indicate that taste sensitivity is influenced by water quality.     

Classification criteria and probability risk maps: limitations and perspectives

Delineation of polluted zones with respect to regulatory standards, accounting at the same time for the uncertainty of the estimated concentrations, relies on classification criteria that can lead to significantly different pollution risk maps, which, in turn, can depend on the regulatory standard itself. This paper reviews four popular classification criteria related to the violation of a probability threshold or a physical threshold, using annual (1996-2000) nitrogen dioxide concentrations from 40 air monitoring stations in Milan. The relative advantages and practical limitations of each criterion are discussed, and it is shown that some of the criteria are more appropriate for the problem at hand and that the choice of the criterion can be supported by the statistical distribution of the data and/or the regulatory standard. Finally, the polluted area is estimated over the different years and concentration thresholds using the appropriate risk maps as an additional source of uncertainty.     

Advanced factor analysis of spatial distributions of PM2.5 in the eastern United States

This work analyzes PM2.5 24-h average concentrations measured every third day at over 300 locations in the eastern United States during 2000. The non-negative factor analytic model, Positive Matrix Factorization, has been enhanced by modeling the dependence of PM2.5 concentrations on temperature, humidity, pressure, ozone concentrations, and wind velocity vectors. The model comprises 12 general factors, augmented by 5 urban-only factors intended to represent excess concentration present in urban locations only. The computed factor components or concentration fields are displayed as concentration maps, one for each factor, showing how much each factor contributes to the average concentration at each location. The factors are also displayed as flux maps that illustrate the spatial movement of PM2.5 aerosol, thus enabling one to pinpoint potential source areas of PM2.5. The quality of the results was investigated by examining how well the model reproduces especially high concentrations of PM2.5 on specific days at specific locations. Delimiting the spatial extent of all such factors that exhibit a clear regional maximum surrounded by an almost-zero outer domain lowered the uncertainty in the computed results.     

Dining at the periodic table: metals concentrations as they relate to recycling

A correlation between the prices of a variety of substances and their dilutions in their initial matrices was shown in 1959 by T.K. Sherwood. The research presented here shows that the relationship holds for engineering metals today, which we termed the metals-specific Sherwood plot. The concentrations of metals in products (e.g., printed wiring boards and automobiles) and waste streams (e.g., municipal solid waste, and construction and demolition debris) were plotted with this correlation. In addition, for the products and waste streams that undergo disassembly at end-of-life, the metals concentrations of the disassembled components were also plotted. It was found that most of the metals that are currently targeted for recycling have post-disassembly concentrations that lie above the metals-specific Sherwood plot (i.e., have concentrations that are more enriched than minimum profitable ore grades). This suggests that material concentration plays a role in the viability of recycling at end-of-life. As products grow in complexity and the variety of materials used, analyses such as this one provide insight for policymakers and those interested in material sustainability into macro-level trends of material use and future recycling practices.     

Evaluation of three gas chromatography-olfactometry methods: comparison of odour intensity-concentration relationships of eight volatile compounds with sensory headspace data

Three gas chromatography–olfactometry methods, i.e. dilution analysis, detection frequency method and posterior intensity method, were evaluated for attribution of odour potency to eight volatile compounds at seven concentration levels. Six serial 1:5 dilutions of a solution consisting of 2-butanone, diacetyl, ethyl acetate, 3-methyl-1-butanol, ethyl butyrate, hexanal, 2-heptanone, α-pinene in pentane were analysed by gas chromatography-olfactometry. In addition, sensory odour intensity of sunflower oil solutions of the compounds at eight concentration levels were determined and related to their headspace concentrations. Posterior intensities correlated reasonably well with sensory odour intensities. Detection frequency data were also highly correlated, whereas dilution analysis led to more diverting results. A large variance among subjects was observed. Therefore, a group of assessors is considered a prerequisite for reliable gas chromatography-olfactometry analysis.     

Relationship between ruminal ammonia and nonprotein nitrogen utilization by ruminants. I. Development of a model for predicting nonprotein nitrogen utilization by cattle.

The influence of ration composition on mean ruminal ammonia concentration was studied by collecting samples of ruminal ingesta from cattle fed rations varying in crude protein and total digestible nutrient content. A minimum of four sampling times distributed throughout the day permitted calculation of mean ruminal ammonia concentrations. Mean ruminal ammonia concentration was positively related to dietary crude protein concentration and negatively related to total digestible nutrient concentration. It is postulated that mean ruminal ammonia concentration may be a useful criterion for predicting efficacy of nonprotein nitrogen supplementation. A quantitative approach for evaluating nonprotein nitrogen supplementation based upon determination of the point at which ruminal ammonia exceeds the requirement (5 mg ammonia nitrogen/100 ml rumen fluid) of the ammonia-utilizing bacteria is proposed. Dietary conditions expected to result in excessive concentrations of ruminal ammonia are defined and recommended upper limits for nonprotein nitrogen supplementation are presented. Theoretical relationships between composition of the unsupplemented ration, amount of nonprotein nitrogen added, and efficiency of nonprotein nitrogen utilization are discussed. The practice of adding nonprotein nitrogen so as to exceed 12 to 13% crude protein in typical dairy or feedlot rations needs to be reevaluated.     

Selection of microalgal growth model for describing specific growth rate-light response using extended information criterion

The effects of light intensity and carbon dioxide (CO2) concentration on the growth of the highly CO2-tolerant green alga Chlorococcum littorale were studied in batch cultures. Four mathematical representations were compared for the specific growth rate-light response curve: a rectangular hyperbolic function, Steele's exponential function, a Poisson function and a hyperbolic tangent function. The hyperbolic tangent function, which is commonly used for representing the photosynthesis-light relationship, gave the best fit as evaluated by the extended information criterion (EIC). EIC proved to be applicable as a criterion to this kind of nonlinear model selection problem. Carbon dioxide, the sole carbon source for photoautotrophic growth of this alga, inhibited the growth rate at concentrations higher than pCO2 of 0.02. A substrate inhibition model was successfully used to simulate the relationship between the specific growth rate and CO2 response.     

Response of Salmonella and Escherichia coli O157:H7 to UV energy

To determine the efficacy of a UV light treatment at 253.7 nm (UVC light) on microbial growth, plates containing tryptic soy agar plus 50 ppm of nalidixic acid (TSAN) were inoculated with known concentrations of five-strain cocktails of Salmonella and Escherichia coli O157:H7 and subjected to different UVC treatments. The concentration of the cocktail inoculum was determined with TSAN prior to inoculation. Serial dilutions were carried out, and inoculation levels of 10(0) to 10(8) CFU/ ml were tested for each pathogen. Multiple replications of doses of UV light ranging from 1.5 to 30 mW/cm2 were applied to different cocktail concentrations, and doses of > 8.4 mW/cm2 resulted in a 5-log reduction of Escherichia coli O157:H7, while a 5-log reduction of Salmonella was observed with doses of > 14.5 mW/cm2. Results for both organisms yielded sigmoidal inactivation curves. UVC light is effective in reducing microbial populations of pathogens on agar surfaces.     

Can a river heal itself? Natural attenuation of metal contamination in river sediment

Sediment sampling of bed sediment from a large river contaminated by mining and smelting was used to determine rates of natural attenuation of metal concentrations. A "natural decay model" was developed from high-resolution temporal data and used to predict when restoration guidelines would be met without restoration and with various degrees of restoration success. The natural decay model estimates that in the most contaminated reaches it will take about 90 years for average concentrations of As, Cd, Cu, Pb, and Zn to fall below "probable effects concentrations" (PEC), i.e. levels above which we expect to see adverse environmental effects. At sites farther downstream, all metals will fall below PEC in <35 ± 8 years. It will take longer to reach "threshold effects concentrations" (TEC), i.e. concentrations at which no effects are expected. But, even in the most contaminated reaches, Cd, Pb, and Zn will reach TEC in <80 ± 57 years, while Cu and As will take ~200 years. Model simulations with different levels of remediation success show that recovery is highly dependent on source reduction and how far the goal is from the basin background concentration. Furthermore, beneficial effects of restoration may be unexpectedly small: for example a likely decrease of ~20% in the source concentration would shorten the time to reach the Cu PEC by only 13 years. We argue that conducting analyses like these can provide insight into remediation approaches and ultimately decrease the cost of restoration by identifying the role of natural attenuation in restoration design and implementation.     

Raoult's Law, Water Activity and Moisture Availability in Solutions

A rationalization of Raoult's law has led to the conclusion that water activity (a_w) is a joint solution property of vapor pressure, solute and solvent concentrations. It is shown from this that a_w is not a measure of the absolute value of the mole fraction of water as indicated by Raoult's law but a measure of only a fraction of the mole fraction of water remaining free in solution available and unbound to solute molecules. The law is shown to overestimate this water activity (a_w) at all dilutions by an amount equal to the product of the mole fraction of solute and the lowered relative vapor pressure the solute generates in solution.     

Identification and quantification of methylglyoxal as the dominant antibacterial constituent of Manuka (Leptospermum scoparium) honeys from New Zealand

The 1,2-dicarbonyl compounds 3-deoxyglucosulose (3-DG), glyoxal (GO), and methylglyoxal (MGO) were measured as the corresponding quinoxalines after derivatization with orthophenylendiamine using RP-HPLC and UV-detection in commercially available honey samples. Whereas for most of the samples values for 3-DG, MGO, and GO were comparable to previously published data, for six samples of New Zealand Manuka (Leptospermum scoparium) honey very high amounts of MGO were found, ranging from 38 to 761 mg/kg, which is up to 100-fold higher compared to conventional honeys. MGO was unambigously identified as the corresponding quinoxaline via photodiodearry detection as well as by means of mass spectroscopy. Antibacterial activity of honey and solutions of 1,2-dicarbonyl towards Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) were analyzed using an agar well diffusion assay. Minimum concentrations needed for inhibition of bacterial growth (minimum inhibitory concentration, MIC) of MGO were 1.1 mM for both types of bacteria. MIC for GO was 6.9 mM (E. coli) or 4.3 mM (S. aureus), respectively. 3-DG showed no inhibition in concentrations up to 60 mM. Whereas most of the honey samples investigated showed no inhibition in dilutions of 80% (v/v with water) or below, the samples of Manuka honey exhibited antibacterial activity when diluted to 15-30%, which corresponded to MGO concentrations of 1.1-1.8 mM. This clearly demonstrates that the pronounced antibacterial activity of New Zealand Manuka honey directly originates from MGO.     

Detection and discrimination of phenol and primary alcohols in water using electronic noses

Electronic nose methodology was used for discriminating between samples of n-primary alcohols and phenol in water. An electronic nose device developed at our laboratory was applied to detect the vapors from the samples. The device consists of 10 nonspecific gas sensors based on tin-dioxide, whose electrical conductivity change when exposed to the vapors. The whole set of responses constitutes a fingerprint associated with the sample for the particular array of sensors. Using this method, we could differentiate among n-primary alcohols, pure phenol, and aqueous dilutions of phenol. Dilutions of primary alcohols and phenol of 1, 3, 5, and 10% (v/v) were assayed. The fingerprints of the dilutions are similar to that of the respective pure (not diluted) alcohol, but with lower signal intensities. Principal component analysis and cluster analysis were performed in order to explore the feasibility of discriminating among the samples. Dilutions of phenol were successfully discriminated from those of primary alcohols. Discrimination of n-octanol and n-decanol from the other alcohols, including phenol, was also achieved. The results suggest that it is possible to discriminate among dilutions of phenol in water and to give an approximate gross estimation of phenol concentration in water.     

Refinements in primary rumen epithelial cell incubation techniques

The objectives of this study were to 1) determine if the number of rumen epithelial cells in primary cell incubation affects the rate of metabolite production, and 2) determine the optimum mode of data expression to standardize reporting criteria. Sections of rumen epithelial tissue were excised from five Holstein heifers and subjected to serial tryptic digestion to isolate cells. Isolated cells had a mean viability of 86% (+/- 1.29) and were incubated at concentrations of 0.5, 1, 5, 10, 20, and 40 million cells per flask. Oxidation of [1-14C]butyrate to 14CO2 and production of acetoacetate (ACAC), beta-hydroxybutyrate (BHBA), lactate, and pyruvate were measured for cell dilution comparisons. Cell number, cell dry matter, cell crude protein, epithelial wet tissue weight, body weight, and metabolic body weight were measured to generate 12 different forms of data expression. Coefficients of variation were calculated for each type of expression. Expressing data per cell number resulted in the lowest variation. Oxidation of [1-14C]butyrate to 14CO2 and pyruvate production per million cells did not significantly differ between treatments for 90-min incubation. Acetoacetate and lactate concentrations were greatest at 0.5 and 1 million cells/flask, respectively, with no differences between 5 to 40 million cells/flask. Production of BHBA for 1 million cells/flask was greater than 0.5 and 40 million cells/flask, but did not change between cell concentrations 5 to 20 million. The BHBA:ACAC concentration ratios for 0.5 and 1 million cell dilutions were both 1.1 to 1 indicating low mitochondrial redox potentials. Concomitantly, lactate:pyruvate ratios for 0.5 and 1 million cells were greater than other cell dilutions, indicating a high cytosolic redox potential. The suggested range of rumen epithelial cells to include in incubations is 5 to 20 million cells/flask. This will minimize experimental error associated with using low cell numbers and the potential for reduced metabolite production caused by incubating large cell quantities. When rumen tissue taken from animals of the same species, size, and stage of development; data adjusted by cell number is preferred. However, it is recommended that cell protein, cell DM, and animal metabolic weight be included to facilitate future comparison between species and laboratories.     

4种柑桔汁的缓冲能力与氮及灰分的二元线性回归关系

果汁含量是果汁的一项重要指标,但目前果汁含量检测方法还不完善,即使有一些检测方法,检测手段也比较繁琐、费时,同时费用较高。对于同一种果汁而言,果汁缓冲能力与果汁的含量呈线性相关关系,但同一类水果不同品种之间缓冲能力是否存在差异以及缓冲能力与水果的那些内在指标密切相关还不清楚。作者选择了南丰蜜桔,砂糖橘、皇帝柑和平远脐橙4种柑桔类水果,制备了相应的柑桔汁,并分别采用检测添加柠檬酸后H+浓度变化的方法检测了其果汁的缓冲系数,采用凯氏定氮法检测了其果汁中氮含量,采用灰化法检测了其灰分质量分数,结果显示这4种柑桔汁缓冲系数与氮含量和灰分呈二元线性相关,回归方程的建立能为利用缓冲能力检测果汁含量的方法提供理论依据。     

The correspondence between U.K. 'action levels' for lead in blood and in water

This paper considers whether the Department of the Environment's water lead concentration criterion for lead pipe replacement and action in individual cases, i.e. 50 micrograms/l in any sample, is too high when set against the Department of Health's advisory action limit for blood lead concentration of 25 micrograms/100 ml. The relationships between blood lead and water lead concentrations found in the Glasgow and Ayr duplicate diet studies, together with unpublished data from Glasgow and Liverpool, indicate that over 10% of people exposed to an average water lead concentration of 100 micrograms/l (the earlier action level) would have blood lead concentrations above 25 micrograms/100 ml, as would about 4% of those exposed to 50 micrograms/l (the Maximum Admissible Concentration in an EEC Directive). For adults, average water lead concentrations should not exceed 30 micrograms/l to ensure compliance with the limit for blood lead, i.e. so that not more than 2% exceed 25 micrograms/100 ml. However, for one of the critical groups, bottle-fed infants (whose diet is 90% water), average water lead concentrations should not exceed 10-15 micrograms/l. The WHO's Provisional Tolerable Weekly Intake (PTWI) for children (25 micrograms/kg body weight) also implies that their water lead concentrations should not exceed 10-15 micrograms/l.     

实时荧光PCR法检测鼠伤寒沙门氏菌

目的 建立实时荧光PCR法检测鼠伤寒沙门氏菌的方法。方法 基于鼠伤寒沙门氏菌II型限制酶基因, 设计引物及Taqman探针, 利用实时荧光PCR进行特异性、灵敏性及模拟样品的检测实验。结果 特异性探针可从25种血清型沙门氏菌(共49株)及11株阴性对照菌株中检测出全部的11株鼠伤寒沙门氏菌。以鼠伤寒沙门氏菌梯度稀释菌液DNA为模板进行实时荧光PCR实验, 菌株模板浓度与Ct值呈良好线性关系, 线性系数(R2)为0.998, 扩增效率90%, 最低检测浓度300 cfu/mL。对已接种鼠伤寒沙门氏菌的4种模拟样品同时进行实时荧光PCR检测和传统方法鉴定, 两者结果一致。结论 此方法特异、灵敏、准确, 适于食品中鼠伤寒沙门氏菌的检测。