Surveillance and Elimination of Bacteriophage Contamination in an Industrial Fermentation Process

Commercial fermentation processes are often vulnerable to bacteriophage due to the lack of genetic diversity and use of high cell density cultures. Bacteriophage infections in these fermentations can have adverse impacts on operability of the production facility and product quality and prevent recovery of valuable bioproducts in the downstream process. Prevention strategies have been developed and optimized through feedback from bacteriophage diagnostic tests, which inform improvements to process design for elimination of entry points, as well as modification of the biocatalyst to reduce or eliminate bacteriophage virulence. In this chapter, we provide case studies for successful elimination of bacteriophage virulence via host modifications, including bacteriophage binding-site modifications on the outer membrane of an Escherichia coli production host, used for commercial manufacture of 1,3-propanediol, as well as application of CRISPR-associated protein 9 (Cas9) for bacteriophage immunity. Finally, we report application of bacteriophage diagnostic methods to fully characterize and eliminate bacteriophage entry points in a commercial fermentation process

Identification of intermediates of in vivo trichloroethylene oxidation by the membrane-associated methane monooxygenase

The rate and products of trichloroethylene (TCE) oxidation by Methylomicrobium album BG8 expressing membrane-associated methane monooxygenase (pMMO) were determined using 14 C radiotracer techniques. [ 14 C]TCE was degraded at a rate of 1.24 nmol (min mg protein) −1 with the initial production of glyoxylate and then formate. Radiolabeled CO 2 was also found after incubating M. album BG8 for 5 h with [ 14 C]TCE. Experiments with purified pMMO from Methylococcus capsulatus Bath showed that TCE could be mineralized to CO 2 by pMMO. Oxygen uptake studies verified that M. album BG8 could oxidize glyoxylate and that pMMO was responsible for the oxidation based on acetylene inactivation studies. Here we propose a pathway of TCE oxidation by pMMO-expressing cells in which TCE is first converted to TCE-epoxide. The epoxide then spontaneously undergoes HCl elimination to form glyoxylate which can be further oxidized by pMMO to formate and CO 2 .Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/74667/1/j.1574-6968.2000.tb09090.x.pd

New Rotation Periods in the Open Cluster NGC 1039 (M 34), and a Derivation of its Gyrochronology Age

Employing photometric rotation periods for solar-type stars in NGC 1039 [M 34], a young, nearby open cluster, we use its mass-dependent rotation period distribution to derive the cluster's age in a distance independent way, i.e., the so-called gyrochronology method. We present an analysis of 55 new rotation periods,using light curves derived from differential photometry, for solar type stars in M 34. We also exploit the results of a recently-completed, standardized, homogeneous BVIc CCD survey of the cluster in order to establish photometric cluster membership and assign B-V colours to each photometric variable. We describe a methodology for establishing the gyrochronology age for an ensemble of solar-type stars. Empirical relations between rotation period, photometric colour and stellar age (gyrochronology) are used to determine the age of M 34. Based on its position in a colour-period diagram, each M 34 member is designated as being either a solid-body rotator (interface or I-star), a differentially rotating star (convective or C-star) or an object which is in some transitory state in between the two (gap or g-star). Fitting the period and photometric colour of each I-sequence star in the cluster, we derive the cluster's mean gyrochronology age. 47/55 of the photometric variables lie along the loci of the cluster main sequence in V/B-V and V/V-I space. We are further able to confirm kinematic membership of the cluster for half of the periodic variables [21/55], employing results from an on-going radial velocity survey of the cluster. For each cluster member identified as an I-sequence object in the colour-period diagram, we derive its individual gyrochronology age, where the mean gyro age of M 34 is found to be 193 +/- 9 Myr, formally consistent (within the errors) with that derived using several distance-dependent, photometric isochrone methods (250 +/- 67 Myr).Comment: accepted for publication in Astronomy & Astrophysic

Presupernova Evolution of Rotating Massive Stars I: Numerical Method and Evolution of the Internal Stellar Structure

The evolution of rotating stars with zero-age main sequence (ZAMS) masses in the range 8 to 25 M_sun is followed through all stages of stable evolution. The initial angular momentum is chosen such that the star's equatorial rotational velocity on the ZAMS ranges from zero to ~ 70 % of break-up. Redistribution of angular momentum and chemical species are then followed as a consequence of rotationally induced circulation and instablities. The effects of the centrifugal force on the stellar structure are included. Uncertain mixing efficiencies are gauged by observations. We find, as noted in previous work, that rotation increases the helium core masses and enriches the stellar envelopes with products of hydrogen burning. We determine, for the first time, the angular momentum distribution in typical presupernova stars along with their detailed chemical structure. Angular momentum loss due to (non-magnetic) stellar winds and the redistribution of angular momentum during core hydrogen burning are of crucial importance for the specific angular momentum of the core. Neglecting magnetic fields, we find angular momentum transport from the core to the envelope to be unimportant after core helium burning. We obtain specific angular momenta for the iron core and overlaying material of 1E16...1E17 erg s. These values are insensitive to the initial angular momentum. They are small enough to avoid triaxial deformations of the iron core before it collapses, but could lead to neutron stars which rotate close to break-up. They are also in the range required for the collapsar model of gamma-ray bursts. The apparent discrepancy with the measured rotation rates of young pulsars is discussed.Comment: 62 pages, including 7 tables and 19 figures. Accepted by Ap

Nutrient Source and Tillage Effects on Maize: II. Yield, Soil Carbon, and Carbon Dioxide Emissions

There is a need to understand the potential benefits of using the biotechnology waste by‐product from manufacturing as a fertilizer replacement in agriculture, by quantifying the economic value for the farmer and measuring the environmental impact. Measuring CO2 emissions can be used to assess environmental impact, including three widely used micrometeorological methodologies: (i) the Bowen Ratio Energy Balance (BREB), (ii) aerodynamic flux‐gradient theory, and (iii) eddy covariance (EC). As a first step in quantifying benefits of applying biotechnology waste in agriculture, a detailed examination of these three methods was conducted to understand their effectiveness in quantifying CO2 emissions for this specific circumstance. The study measured micrometeorological properties over a field planted to maize (Zea mays L. var. indentata ), one plot treated with biotechnology waste applied as a nutrient amendment, and one plot treated with a typical farmer fertilizer practice. Carbon dioxide flux measurements took place over 1 yr, using both BREB and EC systems. The aerodynamic method was used to gap‐fill BREB system measurements, and those flux estimates were compared with estimates produced separately by the aerodynamic and EC methods. All methods found greater emissions over the biotechnology waste application. The aerodynamic method CO2 flux estimates were considerably greater than both the EC and a combined BREB‐aerodynamic approach. During the day, the EC and BREB methods agree. At night, the aerodynamic approach detects and accounts for buildup of CO2 at the surface during stable periods. The BREB systems combined with aerodynamic approaches provide alternate methods to EC in examining micrometeorological properties near the surface

Nutrient Source and Tillage Effects on Maize: I. Micrometeorological Methods for Measuring Carbon Dioxide Emissions

There is a need to understand the potential benefits of using the biotechnology waste by‐product from manufacturing as a fertilizer replacement in agriculture, by quantifying the economic value for the farmer and measuring the environmental impact. Measuring CO2 emissions can be used to assess environmental impact, including three widely used micrometeorological methodologies: (i) the Bowen Ratio Energy Balance (BREB), (ii) aerodynamic flux‐gradient theory, and (iii) eddy covariance (EC). As a first step in quantifying benefits of applying biotechnology waste in agriculture, a detailed examination of these three methods was conducted to understand their effectiveness in quantifying CO2 emissions for this specific circumstance. The study measured micrometeorological properties over a field planted to maize (Zea mays L. var. indentata ), one plot treated with biotechnology waste applied as a nutrient amendment, and one plot treated with a typical farmer fertilizer practice. Carbon dioxide flux measurements took place over 1 yr, using both BREB and EC systems. The aerodynamic method was used to gap‐fill BREB system measurements, and those flux estimates were compared with estimates produced separately by the aerodynamic and EC methods. All methods found greater emissions over the biotechnology waste application. The aerodynamic method CO2 flux estimates were considerably greater than both the EC and a combined BREB‐aerodynamic approach. During the day, the EC and BREB methods agree. At night, the aerodynamic approach detects and accounts for buildup of CO2 at the surface during stable periods. The BREB systems combined with aerodynamic approaches provide alternate methods to EC in examining micrometeorological properties near the surface

Conduction Properties Distinguish Unmyelinated Sympathetic Efferent Fibers and Unmyelinated Primary Afferent Fibers in the Monkey

Different classes of unmyelinated nerve fibers appear to exhibit distinct conductive properties. We sought a criterion based on conduction properties for distinguishing sympathetic efferents and unmyelinated, primary afferents in peripheral nerves.In anesthetized monkey, centrifugal or centripetal recordings were made from single unmyelinated nerve fibers in the peroneal or sural nerve, and electrical stimuli were applied to either the sciatic nerve or the cutaneous nerve endings, respectively. In centrifugal recordings, electrical stimulation at the sympathetic chain and dorsal root was used to determine the fiber's origin. In centrifugal recordings, sympathetic fibers exhibited absolute speeding of conduction to a single pair of electrical stimuli separated by 50 ms; the second action potential was conducted faster (0.61 0.16%) than the first unconditioned action potential. This was never observed in primary afferents. Following 2 Hz stimulation (3 min), activity-dependent slowing of conduction in the sympathetics (8.6 0.5%) was greater than in one afferent group (6.7 0.5%) but substantially less than in a second afferent group (29.4 1.9%). In centripetal recordings, most mechanically-insensitive fibers also exhibited absolute speeding to twin pulse stimulation. The subset that did not show this absolute speeding was responsive to chemical stimuli (histamine, capsaicin) and likely consists of mechanically-insensitive afferents. During repetitive twin pulse stimulation, mechanosensitive afferents developed speeding, and speeding in sympathetic fibers increased.The presence of absolute speeding provides a criterion by which sympathetic efferents can be differentiated from primary afferents. The differences in conduction properties between sympathetics and afferents likely reflect differential expression of voltage-sensitive ion channels

Mars Aeronomy Observer: Report of the Science Working Team

The Mars Aeronomy Observer (MAO) is a candidate follow-on mission to Mars Observer (MO) in the Planetary Observer Program. The four Mariner and two Viking spacecraft sent to Mars between 1965 and 1976 have provided a wealth of information concerning Martian planetology. The Mars Observer, to be launched in 1990, will build on their results by further examining the elemental and mineralogical composition of the surface, the strength and multipolar composition of the planetary magnetic field, the gravitational field and topography, and the circulation of the lower atmosphere. The Mars Aeronomy Observer is intended to address the last major aspects of Martian environment which have yet to be investigated: the upper atmosphere, the ionsphere, and the solar wind interaction region

Estimating global mortality from potentially foodborne diseases: an analysis using vital registration data

<p>Abstract</p> <p>Background</p> <p>Foodborne diseases (FBD) comprise a large part of the global mortality burden, yet the true extent of their impact remains unknown. The present study utilizes multiple regression with the first attempt to use nonhealth variables to predict potentially FBD mortality at the country level.</p> <p>Methods</p> <p>Vital registration (VR) data were used to build a multiple regression model incorporating nonhealth variables in addition to traditionally used health indicators. This model was subsequently used to predict FBD mortality rates for all countries of the World Health Organization classifications AmrA, AmrB, EurA, and EurB.</p> <p>Results</p> <p>Statistical modeling strongly supported the inclusion of nonhealth variables in a multiple regression model as predictors of potentially FBD mortality. Six variables were included in the final model: <it>percent irrigated land, average calorie supply from animal products, meat production in metric tons, adult literacy rate, adult HIV/AIDS prevalence</it>, and <it>percent of deaths under age 5 caused by diarrheal disease</it>. Interestingly, nonhealth variables were not only more robust predictors of mortality than health variables but also remained significant when adding additional health variables into the analysis. Mortality rate predictions from our model ranged from 0.26 deaths per 100,000 (Netherlands) to 15.65 deaths per 100,000 (Honduras). Reported mortality rates of potentially FBD from VR data lie within the 95% prediction interval for the majority of countries (37/39) where comparison was possible.</p> <p>Conclusions</p> <p>Nonhealth variables appear to be strong predictors of potentially FBD mortality at the country level and may be a powerful tool in the effort to estimate the global mortality burden of FBD.</p> <p>Disclaimer</p> <p>The views expressed in this document are solely those of the authors and do not represent the views of the World Health Organization.</p

Conservation agriculture as a climate change mitigation strategy in Zimbabwe

There is a need to quantify agriculture’s potential to sequester carbon (C) to inform global approaches aimed at mitigating climate change effects. Many factors including climate, crop, soil management practices, and soil type can influence the contribution of agriculture to the global carbon cycle. The objective of this study was to investigate the C sequestration potential of conservation agriculture (CA) (defined by minimal soil disturbance, maintaining permanent soil cover, and crop rotations). This study used micrometeorological methods to measure carbon dioxide (CO2) flux from several alternative CA practices in Harare, central Zimbabwe. Micrometeorological methods can detect differences in total CO2 emissions of agricultural management practices; our results show that CA practices produce less CO2 emissions. Over three years of measurement, the mean and standard error (SE) of CO2 emissions for the plot with the most consistent CA practices was 0.564 ± 0.0122 g CO2 m-2 h-1, significantly less than 0.928 ± 0.00859 g CO2 m-2 h-1 for the conventional tillage practice. Overall CA practices of no-till with the use of cover crops produced fewer CO2 emissions than conventional tillage or fallow