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The development of sensitive nucleic acid stains, in combination with flow cytometric techniques, has allowed the identification and enumeration of viruses in aquatic systems. However, the methods used in flow cytometric analyses of viruses have not been consistent to date. A detailed evaluation of a broad range of sample preparations to optimize counts and to promote the consistency of methods used is presented here. The types and concentrations of dyes, fixatives, dilution media, and additives, as well as temperature and length of incubation, dilution factor, and storage conditions were tested. A variety of different viruses, including representatives of phytoplankton viruses, cyanobacteriophages, coliphages, marine bacteriophages, and natural mixed marine virus communities were examined. The conditions that produced optimal counting results were fixation with glutaraldehyde (0.5% final concentration, 15 to 30 min), freezing in liquid nitrogen, and storage at −80°C. Upon thawing, samples should be diluted in Tris-EDTA buffer (pH 8), stained with SYBR Green I (a 5 × 10−5 dilution of commercial stock), incubated for 10 min in the dark at 80°C, and cooled for 5 min prior to analysis. The results from examinations of storage conditions clearly demonstrated the importance of low storage temperatures (at least −80°C) to prevent strong decreases (occasionally 50 to 80% of the total) in measured total virus abundance with time.
It has been well established that viruses are abundant and important components in aquatic ecosystems (1, 18, 32, 41, 44) since they have been shown to be a major source of mortality for bacteria and eukaryotes (4, 6, 31, 36, 38, 40). Because of their relatively short infection cycles, virus populations are highly dynamic and result in rapid changes in both total numerical abundance and diversity (5, 11, 13-15, 24, 33-35, 37, 43, 45). Assays for rapidly counting viruses with high precision are, therefore, beneficial for studies of viral ecology in the laboratory as well as the field.
Viruses have traditionally been enumerated by culture-based methods (e.g., plaque counts and most-probable-number assays) and transmission electron microscopy (1, 3, 15, 39). These techniques were either selective for viruses infectious for a specific host or very time-consuming. The introduction of high-fluorescence-yield nucleic-acid-specific stains in combination with epifluorescence microscopy (19, 20, 30) significantly improved the quantitation of viruses. With the recent introduction of flow cytometric detection and enumeration of free viruses (7, 27, 29), speed of analysis and accuracy of counting was further improved. This method no longer relies on the skills of the operator. Direct comparison showed that epifluorescence- and flow cytometry-based virus counts were highly comparable (27). Brussaard et al. (7) have shown that a variety of viruses of different morphologies and genome sizes could be detected by flow cytometry. In combination with SYBR Green I as the fluorescent stain, flow cytometry has been used successfully to count viruses from laboratory experiments (8, 9) and in natural samples (11, 12, 24, 25, 27, 43).
The methods used for flow cytometric analyses of viruses in natural samples have, however, not been consistent (7, 12, 29, 43). The few reports on methodologies suggest that optimal detection of virus particles depends on various factors. For example, the study by Marie et al. (27) indicated that the type and concentration of fluorescent stain, the solution used to dilute the sample, the incubation temperature, and the addition of a detergent may all influence the quality of the fluorescent signal. Chen et al. (12) tested the use of the fluorescent dye SYBR Gold for viruses infecting the cyanobacterium Synechococcus spp. Analyses of unfrozen samples showed that staining with SYBR Gold resulted in a higher fluorescence than SYBR Green I. Nevertheless, Marie et al. (27) reported that samples frozen in liquid nitrogen gave better results than unfrozen samples. There are no data available yet on the use of SYBR Gold in combination with freezing. Nor are there currently studies published reviewing the methods used in flow cytometric analyses of viruses without the use of antibodies.
In order to optimize the flow cytometric detection and enumeration of free viruses, a detailed evaluation of a broad range of factors (storage conditions and staining specifications) that potentially affect the quality of the fluorescent signal and subsequently the counting efficiency is presented here. Tests were performed on a variety of different viruses, including four representatives of marine phytoplankton viruses, one freshwater phytoplankton virus, one virus infecting a cyanobacterium, four characterized coliphages, three uncharacterized marine heterotrophic bacteriophages, and two natural mixed marine virus communities.
Virus strains.The viruses CeV-01B, MpVUF10-38, PoV-01B, and PpV-01B, which infect the photosynthetic unicellular organisms Chrysochromulina ericina, Micromonas pusilla, Pyramimonas orientalis, and Phaeocystis pouchetii were kindly provided by G. Bratbak (University of Bergen, Bergen, Norway); PBCV-1, which infects Chlorella sp., was provided by J. Van Etten (University of Nebraska); and S-PMS2, which infects Synechococcus sp., was provided by W. Wilson (Marine Biological Association, Plymouth, United Kingdom). The coliphages Lambda, T2, T4, and T7, which infect the heterotrophic bacterium Escherichia coli were kindly provided by G. Bratbak (culture collection of the Department of Microbiology, University of Bergen). The marine bacteriophages T-φHSIC A, φ16, and T-φD1B, which infect Listonella pelagia, Vibrio parahaemolyticus strain 16, and Flavobacterium sp., respectively (22, 23, 42), were kindly provided by J. Paul (University of South Florida). Natural seawater samples collected from Dutch coastal waters (NS1) and the southern North Sea (NS2) in autumn were also analyzed.
Treatments.Samples were subjected to different treatments (Table 1), including type of fixative and percentage fixative used, storage temperature and length of storage, type and concentration of the stain and dilution solution, pH of the Tris-EDTA (TE) buffer, length and temperature of incubation, addition of detergents, and dilution factor. The treatment used as a reference was fixation with glutaraldehyde (0.5% final concentration, EM-grade; Merck) for 30 min at 4°C, followed by freezing in liquid nitrogen (N2; −196°C) and storage at −80°C (7, 27). When thawed at 35°C for a few minutes (samples should still be cool), samples were diluted in TE buffer (10 mM Tris-HCl and 1 mM EDTA, pH 8) and incubated with SYBR Green I (5 × 10−5 dilution of commercial stock; Molecular Probes) at 80°C for 10 min in the dark.
Two fixatives, glutaraldehyde and formaldehyde, were tested at different concentrations. Dilution solutions tested were Tris (pH 8), TE buffer (pH 7 to 8.9), distilled water, phosphate-buffered saline buffer, and sterile seawater (0.2 μm filtered). Fluorescent stains tested were SYBR Green I and SYBR Gold (Molecular Probes). To mimic the effect of dilution of seawater virus samples in TE buffer, sterile filtered seawater was added at different final concentration. The effect of citrate and various detergents (Triton X-100, Tween 80, NP-40, and sodium dodecyl sulfate [SDS]) was also tested.
Sample analysis.Samples were analyzed by using a FACSCalibur flow cytometer (Becton Dickinson, San Jose, Calif.) equipped with a 15-mW 488-nm air-cooled argon-ion laser and a standard filter setup. The trigger was set on green fluorescence. To avoid coincidence of viral particles (i.e., two or more particles being simultaneously within the sensing zone), the samples were diluted such that the event rate was between 100 and 1,000 viruses s−1. Yellow-green fluorescent latex microspheres (0.85 μm in diameter; Polysciences, Inc., Warrington, Pa.) were added as an internal reference to all samples. Readings were collected in logarithmic mode (at least 5,000 events per sample) and analyzed with CYTOWIN (freely available at http://www.sb-roscoff.fr/phyto/cyto.html ). The data were converted to linear values and normalized to the bead internal standard. Green fluorescence (GFL), total counts, and side scatter (SSC) were recorded. Because viruses are generally smaller than the wavelength of the laser light used (7), the SSC signal cannot be used as an indicator of size and granularity but is very useful as a discriminator of the virus populations (7, 21).
Differences in cytograms (plotting GFL versus SSC) for the different viruses tested are shown in Fig. 1. The windows indicated were used to discriminate the virus of interest and to determine the total virus count and examine specific staining characteristics. Total virus counts were obtained by correcting the measured total counts for noise (typically having the lowest GFL) with sterile and filtered (0.2-μm pore size) seawater or medium as blank. Tests with virus-free lysate (filtered to <30 kDa) showed results comparable to tests with sterile and filtered seawater or medium.
The marine bacteriophage yielded low fluorescence signals (Fig. 1) at the normal settings of the green fluorescence photomultiplier. Increasing the voltage of the photomultiplier improved the results slightly but simultaneously increased the noise. Consequently, there was no real improvement in the signal-to-noise ratio.
Statistical analyses of different treatments were performed by using Systat 10 software. To fulfill the assumptions of analysis of variance, all data were log transformed. Either a two-way analysis of variance (ANOVA) with viruses as a random factor (block) and treatment on 2 to 13 levels or a four- or five-way ANOVAs with interaction terms and two levels for each treatment was used. A probability of P < 0.05 was taken to conclude that the treatment differed significantly in its effect on the measured value.
The standard deviations for GFL, total counts, and SSC among replicates (n = 5) were <5% for the phytoplankton virus (PpV) and <12% for bacteriophages Lambda and φ16. These results show that a detailed comparison of staining characteristics and measured abundance for the different treatments and viruses is valid. Furthermore, combining four different viruses (CeV, MpV, PpV, and S-PMS2) in one sample provided identical results compared to the analysis of individual viruses. This indicates that the results obtained with individual viruses can be interpolated to mixed-virus situations such as those found in nature.
The statistical variance analyses of the different treatments are summarized in Table 2 and 3. The analysis of four treatments (freezing, fixation, heating, and addition of detergent) for all viruses in the present study indicates that there were significant differences for total virus counts, GFL, and SSC signals between the group consisting of phytoplankton viruses and the group consisting of heterotrophic bacteriophages (Table 2). Therefore, these two virus groups were also analyzed separately in order to obtain insight into the factors and extent to which the groups differed.
Fixatives.Fixation had a significant positive effect on GFL and SSC compared to that of unfixed samples (Table 2). A more detailed statistical analysis showed that the improved GFL signal was due to the heterotrophic bacteriophages and that the improved SSC signal was due to the phytoplankton viruses (Table 2). The extent to which the viruses were positively affected by fixation depended on the type of virus (interaction effect). There was no difference in signal for up to 1 h of fixation (0.25, 0.5, 1, 3, and 18 h tested), and no significant difference between fixation at room temperature and 4°C was recorded. The type of fixative (glutaraldehyde or formaldehyde), as well as the final concentration of fixative (0.1 to 2% for glutaraldehyde and 0.4 to 4% for formaldehyde) were tested (Table 3). The use of formaldehyde rather than glutaraldehyde occasionally led to reduced virus counts (for example, the marine phages T-φHSIC showed a 20% reduction). The total counts were slightly reduced at concentrations of formaldehyde of >0.4% (reductions typically <10%, but a 55% reduction was recorded for T-φHSIC A). Although no significant trend was observed for the different concentrations of glutaraldehyde, reductions in virus abundance were occasionally observed at concentrations of >1% (e.g., S-PMS2 and Lambda).
Incubation temperature.Heating of the samples to 80°C prior to analysis significantly increased the total virus count (Table 2) and improved the staining characteristics (GFL and SSC). Measured virus abundance was most enhanced for the group of heterotrophic bacteriophages (up to eightfold for φ16 and T-φD1B). Heating showed significant interactions with the type of virus and fixation (Table 2). Five different temperatures in the range of 20 to 90°C were tested in order to find the optimal temperature for staining of virus samples (Table 3). In general, heating the samples to 80 to 90°C showed the best results. Analysis of the length of the incubation period (1, 4, 7, 10, 13, 16, and 19 min) showed a significant effect for GFL (Table 3), with the signal increasing until 10 min.
Dyes.The concentration of SYBR Green I (ranging in dilution from 5 × 10−5 to 25 × 10−5 of the commercial stock) significantly affected total virus counts (Table 2B), showing 5 to 45% reductions in measured virus abundance when 1 × 10−4 to 25 × 10−5 of the commercial stock was stained compared to the optimal dilution of 5 × 10−5 of the commercial stock. The addition of SYBR Green I after the samples were heated (instead of prior to heating) negatively affected the GFL signal and the total counts for the viruses that infect heterotrophic bacteria. For example, marine bacteriophages φ16 and T-φD1B showed a 50 to 70% reduction in GFL and a 40 to 50% reduction in counts.
Staining with another sensitive nucleic acid-specific stain, SYBR Gold, resulted in slightly (but significantly) lower GFL signals of the stained viruses (Table 3). No significant effect was recorded for the total virus counts when all tested viruses were taken into account, but for some of the individual viruses and a natural sample, staining with SYBR Gold resulted in reduced total counts compared to staining with SYBR Green I (Fig. 2). The GFL signal decreased with increasing concentrations of dye (from 5 × 10−5 to 25 × 10−5 of the commercial stock; P < 0.05, Table 3). To allow comparison with published work on cyanophages using SYBR Gold, the virus S-PMS2 (infecting Synechococcus sp.) was specifically tested in more detail (Table 4). Staining with SYBR Gold, compared to staining with SYBR Green I, resulted in significantly lower total counts. At dilutions of >5 × 10−5 of the commercial stock, both the total virus count and the GFL signal were further reduced (Table 3). Heating the sample affected the total counts, as well as GFL and SSC signals, positively. Freezing the samples in liquid nitrogen prior to analysis also had a significant positive effect on the measured total virus abundance.
Dilution solution.To avoid coincidence during flow cytometric analysis, the virus samples were normally diluted. The choice of solution (seawater, Tris buffer, TE buffer, phosphate-buffered saline buffer, or distilled water) had a significant effect on the measured virus abundance (Table 3), showing highest counts when diluted in TE or Tris buffer. The lowest counts were obtained when the virus sample was diluted in distilled water or seawater. Dilution in TE buffer also yielded the best GFL signal, while the pH of the TE buffer (7, 7.4, 7.8, 8, 8.4, or 8.85) had no significant effect (Table 3), although individual viruses reacted very differently (e.g., PpV showed 40% reduction in total count at a pH of <8).
Additives.The ratio of TE buffer to seawater is influenced by the dilution factor of the marine virus sample. To simulate the effect of various percentages of seawater samples at different dilutions, sterile and filtered (0.2-μm pore size) seawater at final concentrations of 1, 2, 5, 10, 20, and 50% (vol/vol) was added to highly diluted virus samples (>1,000 times diluted in TE at pH 8). With increasing concentrations of seawater, a significant decline in GFL signal and in total counts was recorded (Table 3). To test whether these effects were due to the high salt concentration, sodium chloride (NaCl) was added to the samples at final concentrations of 2, 5, and 10% (vol/vol). No significant effects were recorded as a result of the NaCl additions.
The addition of Triton X-100 (0.1% [vol/vol]) had no significant effect on total virus counts or staining characteristics (Table 2) but was occasionally found to improve the coefficient of variance of the GFL signal. It was hypothesized that detergents would facilitate the permeabilization of the viral particles and would therefore enhance the GFL signal upon staining. A significant effect on the total virus counts was recorded (Table 3) by using Triton X-100, Tween 80, NP-40, and SDS (all at 0.1% [vol/vol] final concentrations). However, there was no significant effect when the anionic surfactant SDS, negatively affecting the abundance of the heterotrophic bacteriophages (reductions up to 95%), was omitted from the statistical analysis. The problem with the addition of surfactants was the generation of high levels of noise. As long as the viruses have GFL signals well above the level of the produced noise (for example, many of the phytoplankton viruses), this will not be bothersome. However, it could present a problem when virus samples with relative low GFL levels are analyzed. Sufficient blanks should be taken to allow subtraction of the noise from the total virus counts in the samples.
Another factor that was hypothesized to affect the discrimination and enumeration of viruses by flow cytometry is citrate. No significant effect of citrate addition (final concentrations of 1, 2, 5, 10, 20, or 50 mM) was recorded in the present study (Table 3).
Storage.A range of temperatures—4, −20, −80, and −196°C (deep freezing in liquid nitrogen)—for storage was tested for 7 of the 12 viruses studied. Storage temperature significantly affected total virus counts (Table 3), although large differences in total count were recorded among the different viruses tested (Fig. 3). The optimal storage protocol was deep freezing in liquid nitrogen (followed by storage at −80°C for convenience). The present study clearly demonstrates that storage of virus samples should be conducted at very low temperatures (Table 3, Fig. 3 and 4). Some of the samples stored at 4°C for a few hours resulted in only ca. 10% of the measured abundance compared to samples deep frozen in liquid nitrogen and stored at −80°C (Fig. 3A). Figure 3B shows that 1 month of storage at 4 or −20°C led to high reductions in total virus counts (down to a few percent of the total count at t = 0). Also, the GFL signal was significantly reduced for the samples stored at 4°C (Table 3). Because it is not uncommon to store samples at 4°C before analysis, a separate statistical analysis was performed to test for variance in the length of storage time at this temperature. Total counts and SSC signal were negatively affected by storage duration (Table 3), showing reductions in total virus counts of 80% within 14 days (Fig. 4).
The development of sensitive nucleic acid stains, in combination with flow cytometric techniques has allowed identification and enumeration of very small particles such as viruses in aquatic systems (7, 8, 11, 12, 24, 25, 27, 29, 43). So far, however, studies using flow cytometry to count viruses in aquatic ecosystems have not used consistent methods. The present study was executed to optimize the flow cytometric enumeration of viruses and promote consistency of the methodology used. Total virus counts, green fluorescence (GFL) and side scatter (SSC) of the viral particles were recorded for a variety of different viruses (including representatives of phytoplankton viruses, cyanobacteriophages, coliphages, marine heterotrophic bacteriophages, and natural mixed marine virus communities) that where subjected to a range of different treatments potentially influencing detection and quantitation of virus particles.
Virus abundance.In general, total virus counts were significantly and positively affected when samples were frozen in liquid nitrogen, heated at 80°C (10 min, followed by 5 min cooling down of the sample), and diluted in TE-buffer. It is recommended to add the fluorescent dye prior to heating of the sample, because staining of the heterotrophic bacteriophages was less intense when the fluorescent dye was added after heating. Marie et al. (27) did not find any effect of heating and time of addition of the dye, however this study performed the analyses with a virus infecting the phytoplankter Phaeocystis pouchetii (i.e., PpV). Consistent with these results, in the present study there was no increased counting efficiency detected upon heating for this virus. With the majority of the viruses in marine environments being bacteriophages, and two of the tested marine heterotrophic phages showing significant reduction in total virus counts at 60°C (12 to 65% reduction compared to sample heated at 80°C), even a reduction in temperature to 65°C as used by Evans et al. (17) can be expected to underestimate the total natural virus concentration.
Natural samples normally have to be diluted before flow cytometric analysis in order to avoid coincidence. Dilution in TE-buffer (pH 8) gave the best results for virus enumeration in conjunction with staining quality, as was also demonstrated by Marie et al. (27). Dilution in seawater (0.2-μm pore size filtered and autoclaved) should be avoided because of the reduction in total virus counts and a reduction in the staining signal. Since counting efficiency was reduced with increasing percentages of seawater, the dilution factor of marine virus samples in TE-buffer should be as high as possible (preferably at least > 50) while keeping statistically significant virus counts. In the present study there was a 40% reduction in estimated viral abundance for the marine heterotrophic bacteriophage and natural samples diluted 2-fold (50% final concentration of seawater). Measured virus abundance was still 15% reduced when those samples were 10-fold diluted. Studies using low dilution factors for virus samples (17) can thus be expected to underestimate the total virus concentration. Although a high dilution factor reduces the negative effect of seawater, the statistically optimal event rate should not become too low (optimal between 100 and 800 per s [27, 29]; unpublished results using different virus samples). In case the virus event rate becomes too low, one has to switch to a higher flow rate.
Staining the viruses with the nucleic-acid-specific dye SYBR Gold did not significantly enhance total virus counts compared to SYBR Green I, and in fact some individual virus samples, such as the natural sample NS 2, showed lower total counts when stained with SYBR Gold. Chen et al. (12) used SYBR Gold at a dilution of 2.5 × 10−4 the commercial stock to stain viruses (including viruses infecting Synechococcus sp.). The present study shows that at this concentration the measured virus abundance is significantly reduced compared to a dilution of 5 × 10−5 the commercial stock. This concentration is also optimal for SYBR Green I and the use of relatively high concentrations of dye is therefore not suggested for routine use. In addition to counting performance, virus staining characteristics with SYBR Green I were significantly better than with SYBR Gold, indicating the preference of using SYBR Green I for standard analysis. Other fluorescent dyes tested (7, 27) have not been shown to improve staining quality nor estimated total virus counts.
Chen et al. (12) used a high concentration of glutaraldehyde (2.5% final concentration) to fix virus samples (natural as well as viruses infecting Synechococcus sp.). This is not recommended as it was found to reduce the total cyanophage count in the present study. The phytoplankton virus abundances were lower when fixed with glutaraldehyde at final concentrations > 0.5%. The tests with glutaraldehyde versus formaldehyde in the present study were not conclusive, but the natural seawater samples showed higher GFL signals when fixation was performed with glutaraldehyde. This will allow better separation of viruses with a relative low green fluorescence upon staining. Formaldehyde is generally used in studies using epifluorescence to enumerate viruses (2, 20, 30), because it has a lower background noise on the 0.02 μm pore-sized Anodisc filters. The use of flow cytometry, however, does not restrict one to formaldehyde fixation. Because fixation with glutaraldehyde is a slower process than with formaldehyde, time should be given to fix the viral nucleic acids effectively (10). A fixation time of 15 to 60 min was found to be most efficient in the present study, as total virus counts began to decline after fixation for several hours, even at 4°C. Long fixation (12 h) of the virus samples before freezing, as is described in the study by Evans et al. (17), is therefore not advised.
The temperature at which the virus samples were stored postfixation was found to be critical. Nonfrozen samples showed a rapid loss of counts, and thus storage at 4°C is strongly discouraged. Studies using nonfrozen fixed virus samples can be expected to underestimate the total virus concentration (12), with the extent depending on the length of time at 4°C (frequently, total virus counts were down to 20% within 14 days storage). A few other studies have reported reductions of viral abundances in fixed samples upon storage at 4°C (16, 27, 46), but a comparison of storage temperatures has not been tested before. When comparing total virus counts of different studies, it is important to take the type of storage into account. Actually, only the samples that were frozen in liquid nitrogen upon fixation (whether stored at −80°C or not) did not show significant declines in total virus numbers over time (tested up to 6 months). For optimal results and to standardize methodology it is recommended for future studies to freeze virus samples in liquid nitrogen.
Staining characteristics.Working close to the limits of staining methodology and the technical performance of a flow cytometer, the level of green fluorescence (GFL) is crucial for optimizing the detection of free viruses (7). The GFL signals of the stained viruses were positively affected by heating the samples prior to analysis, and by incubation time (optimal signal after 10 min). It is hypothesized that heat treatment affects the permeability of the viral capsid and denatures the nucleic acids (at least partly), thereby improving the staining characteristics of the dye. Fixation improved the staining of the heterotrophic bacteriophages, presumably the permeabilized membranes of the virus particles provided better access for the dye.
Additives such as citrate have been reported to improve both the staining quality and the stability of the SYBR Green I fluorescence when used for cell cycle analysis of phytoplankton (28). This could not be confirmed for viruses (present study). Addition of detergents, which potentially make the virus capsid permeable and thus allowing easier penetration of the dye into the cells (26, 27), did not significantly improve of the staining characteristics either. To the contrary, high levels of noise were generated with the addition of detergents. The use of Triton X-100 for the analysis of natural samples (12, 17), typically containing high percentages of low-staining bacteriophages, is therefore not recommended. When working with controlled mixed systems with relative high staining signals, however, the addition of detergent may improve the separation of the different virus populations.
Evaluating the effects of a broad range of variables on the staining specifications of aquatic viruses shows that there can be large variations between different viruses. Intrinsic features such as the packaging degree of the nucleic acids, and the GC content of the viral genome can be expected to affect the entry and binding of the fluorescent dye, and help to explain the different responses to the methodological variables. Whether the phage genome was circular (Lambda and T4) or linear (T7), appeared unimportant for staining characteristics.
Conclusions.When studying a specific virus type, it is best to determine the combination of variables that results in optimal staining features and counting performance. In the field, environmental parameters may potentially affect the staining or counting efficiency and so it is recommended that investigators document the best protocol for their specific system under study (sampling location). Since this will frequently not be possible, the most practical solution is to be consistent in the methodology used to enumerate viruses. The present evaluation of sample treatments, using not only various specific cultured phytoplankton viruses and heterotrophic bacteriophages but also natural marine samples, indicates that one specific set of variables provided the best results for flow cytometric detection and enumeration (Fig. 5). In summary, virus samples are best fixed with glutaraldehyde (0.5% final concentration, 15 to 30 min at 4°C), frozen in liquid nitrogen, stored below −80°C, diluted in TE buffer (pH 8), stained with SYBR Green I at a final dilution of 5 × 10−5 the commercial stock, and incubated for 10 min in the dark at 80°C (allow cooling down of sample for 5 min prior to analysis).
This study was supported by the European Commission through the research programs Training and Mobility of Researchers (MAST3.CT96-5033[DG12-ASAL]) and Environment and Sustainable Development (EVK3-CT-1999-00015 BIOHAB).
I thank G. Bratbak, J. Paul, C. Suttle, J. Van Etten, and W. Wilson for providing the viruses used in this study. I also thank Harry Witte for help during the final stages of the data analysis and Gerhard Herndl and two anonymous reviewers for constructive comments on the manuscript.
- Received 24 March 2003.
- Accepted 25 November 2003.
- Copyright © 2004 American Society for Microbiology


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text: Trusty and welbeloved we greet you well. Whereas we have taken into our Royal consideration the Loyalty Integrity and Ability of our trusty and welbeloved Thomas Child Esqre We have thought fit hereby to authorize and require you forthwith to cause Letters Patents to be passed under our seal of that our Province of North Carolina in America for constituting and appointing him the said Thomas Child our Attorney General of and in our said Province the said Office being vacant by the promotion of John Montgomery Esqre late Attorney General there deceased to the Office of Chief Justice in our said Province to have hold exercise and enjoy the said office of our Attorney General unto him the said Thos Child during our Pleasure and his residence within our said Province together with all and singular the Rights Salaries Fees Profits Privileges and Emoluments thereunto belonging or appertaining and for so doing this shall be your Warrant and so we bid you farewell. Given at our Court at St James's the 28th Day of Februry 1745-6. in the nineteenth year of our reign.


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This article made me wonder if the theme of "honor and shame" in the Mediterranean plays out in planes other than the family, morality, and community, but also in food. On page 63, Meneley says, "I was told by several Italian olive oil producers that if an oil does not achieve an extravirgin destination, they do not bottle it under their own label, metaphorically disowning the 'impure' daughter." I found this very interesting because while at first glance it may not seem very different that what many American companies may do, the language used implies otherwise. It is likened to a daughter and the word "disowned" is even used. In America, we are used to food companies selling fairly low-quality food as high-quality food, and using clever advertising to make the food seem enticing for reasons other than quality. Many Italian olive oil companies, on the other hand, do not seem to be so quick to distract from the quality. Rather, companies like the ones mentioned in the article are so unwilling to sacrifice quality and extravirginity that they will not sell less pure olive oil under their own name. This, to me, implies that pureness of not only daughters, but olive oil too, is a basis for honor and shame.
Author: at340215
Writing Women's Worlds, ch. 1 & 3
Lila Abu-Lughod writes in a way that reflects the group she studies. She mentioned early on that the people of the Bedouin community had a tendency to describe the past not in terms of a personal journey that they had undertaken, but in terms of dramatic events and detailed memories. So, she writes her ethnography in this style.
The entire first chapter is the story of Migdim's life experiences, which reveal the patrilineality in her community. It is composed mainly of long quotes and detailed descriptions, which tell of how Migdim evaded and got out of potential marriages, the time she finally married, and the time she lost her husband.
Migdim gained some control over the events in her life by sabotaging herself. She needed to go crazy if she wanted to get out of a marriage arrangement. First, her father tried to marry her off to one of her paternal first cousins, who she knew and did not like. She poured the dye that was prepared for her tent all over herself, tried to ask spirits to possess her, and did not eat for nearly two weeks.
After evading multiple marriage arrangements, Migdim finally married Jawwad and had children with him: four boys and three girls. When a woman gives birth to a girl, it is different than if she gives birth to a boy. Lila witnessed a few of the women arguing over whether having a girl is sad or whether it is a happy thing. The thing is, the girls are not truly the mother's. They leave when they get married, and it is difficult for a mother to emotionally prepare for that (for the daughter leaving at any time). Sons are good to have because they stay. If a woman's husband dies, she is not alone, although the sons may very well neglect her when they marry.
Before the group settled into houses, things were different. Migdim remembers how women used to dress more modestly. They wore "heavy black shawls" rather than "flimsy pieces of cloth". On the other hand, weddings did not used to be sex-segregated. Young men and women used to sing and talk at weddings, but now Migdim says weddings have lost their appeal.
The third chapter goes back to the themes touched on in the first chapter: marriage, attitudes about having girls versus boys, and midwives.
It is interesting to me that circumcisions were celebrated within the Bedouin community. What's more, sometimes those circumcisions are celebrated at weddings. This is done if the hosting family wants guests to only have to go to one event instead of two and to only give one gift instead of two. I find this juxtaposition of events strange, as it seems to suggest that circumcisions and weddings are of equal or at least similar importance. To the Bedouin community though, a circumcision celebrated with a wedding may not even seem like a juxtaposition at all.
Sagr, who is a father, tells Lila that girls have less strength of will, yet are more dear to fathers than are sons. He also says that fathers need to test their son a lot to make sure he turns out well, but for daughters, fathers need only "make sure she marries well and can take care of herself." Two things he says that are very telling of his culture are, "as long as her father is alive, a girl does not feel any pain" and the proverb, "it is possible for a child to abandon his mother or father, but not the reverse" (page 160).
My question is, what does Lila Abu-Lughod's writing style in Writing Women's Worlds reveal about the Bedouin community which she studied?


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Perverse role of US think-tanks
February 25, 2010 1 Comment
Asif Haroon Raja
There are 1777 think tanks based in USA. Likewise, large numbers are working in western countries. These think tanks as well as leading newspapers have remained focused on justifying invasion of Afghanistan and Iraq; to cover up crimes committed by US-NATO forces; to paint radical Muslims within targeted countries as human eating monsters and US forces as liberators. Rather than postulating how to defuse the explosive situation which is threatening world peace, their pens never dry up spewing out poison to further stoke fires of chaos and anarchy. Their write ups are based on tutored themes given by vested interests aimed at distorting facts, spreading disinformation, uncertainty and paranoia so as to keep the pot of war on terror boiling.
They portray the aggressors as virtuous free of any faults and victims of aggression as evil. Great majority of think tanks controlled by Jews are intensely prejudiced, irrational and imbalanced. They work on a given agenda to achieve sinister objectives. They have played a major role in demonizing Islam and Muslims and creating disharmony between faiths. 9/11 was over played to turn the guns of the west against the Muslims. Terrorism was made into a buzzword and every Muslim sporting a beard was suspected to be a terrorist and hounded. New laws were framed to humiliate and persecute Islamists. All freedom struggles waged by Muslims were categorized as terrorism. None discussed the root causes of terrorism or defined terrorism, or state terrorism objectively. Think tanks converted Al-Qaeda into a Frankenstein monster, which haunt policy makers in Washington, London, Tel Aviv and New Delhi. Like Osama bin Laden led Al-Qaeda, Mulla Omar led Afghan Taliban were also described as fiends. All these militants whom they now profusely hate were the blued-eyed of USA and western world in the 1980s since it were these holy warriors, now turned into terrorists, who bestowed the status of sole super power upon USA. Without their heroics USA could never ever have fulfilled its dream. As long as the US loved the Mujahideen they were loved by the free world. No sooner they began to hate them and became egotistical, unilateral and self righteous; it triggered anti-Americanism and the whole world began to hate USA for being selfish, untrustworthy and arrogant.
Judging from the track record and conduct of these jaundiced think tanks, it is quite evident that they are hell-bent to keep USA and Muslim world on a warpath so as to kill two birds with one stone. Clash of civilization between Christianity and Islam suit the agenda of Israel and India, both aspiring to become world powers. The Zionists had first pitched US led western world and Muslim world against Communism and after the fall of latter in 1991, Islam became their next target. Among the Muslims those with religious bent of mind and closer to religion were marked for annihilation with the help of secular Muslim leadership and backed by ultra liberal Muslims. American Jews played a key role in coloring the perceptions of US leaders. In order to turn American and western public hostile against Muslims, 9/11 was master minded by the Jews which changed the dynamics of the globe. Their grisly role has now been sufficiently exposed.
When George W. Bush decided to invade Afghanistan to avenge terrorist attacks allegedly master minded by Osama bin Laden and to ensure homeland security, India offered its full services and tried hard to convince Washington to tackle both Afghanistan and Pakistan simultaneously. Bush Administration ignored the counsel and preferred Pakistan over India because of its remarkable role in war in Afghanistan against Soviet forces in 1980s. As long as Washington trusted Pakistan and its premier institutions it made good progress. No sooner it started to mistrust Islamabad and indulged in undesirable 'do more mantra' under the influence of India and Israel and US think tanks, it lost direction and things began to go haywire and once the initiative slipped out of US hands it could not be regained.
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text: Our 2013 S85 has a high pitch whine at low speeds whenever the accelerator is depressed. The sound fades as the speed increases, but it annoyingly announces my arrival every day at work and later when I arrive home. So much for Tesla's silence. Several months ago the Scottsdale Service Center took the car for a test drive and acknowledged the sound as a "known issue" and indicated that "engineers are working on a fix that has not yet been released." The service rep indicated that the car was safe and the sound was not an indicator of a potential problem with the drivetrain. I was told that I would be contacted when a solution become available. I returned to the service center last week to report that the sound was getting worse and was advised again that no solution was yet available and the service rep didn't know when one might become available. When I again asked if this was a potential indicator of a developing problem with the drivetrain, this time there was no reassurance that the sound was non-problematic in terms of the potential for a breakdown. I wonder how many of us have this problem and if other service centers have been authorized to fix the annoying noise.


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Ian Boles, B.A., M.A.
Email: email@example.com
Phone: (714) 484-7185
Office Hours: Meetings accommodated upon request.
Location: Geography Dept., Rm. 238, Humanities
About Professor Boles
Greetings! My name is Ian Boles. I am an adjunct professor of geography here at Cypress College. My main academic interests are Geographic Information Systems (GIS), world geography, and population geography. I encourage you to explore our course offerings in the outstanding Department of Geography at Cypress College.
In my day job, I am the GIS Analyst at the Center for Demographic Research based on the campus of California State University Fullerton.
Like any good geographer, I have a love affair with travel. Whether it is a weekend trip to the Sierras or a trip abroad, I make it a point to explore our planet with all of my senses, and I encourage my students to do the same.
I have traveled to China, Cambodia, Thailand, Indonesia, Singapore, Australia, New Zealand, Iceland, France, the United Kingdom, the Netherlands, Belgium, Germany, Italy, Switzerland, Austria, Czech Republic, Monaco, Turkey, Costa Rica, Nicaragua, El Salvador, Jamaica, Mexico, Canada, the great states of Alaska and Hawai'i, and across the greatest state of all, California.
Recommended Course
GEOG 230 C: Intro to GIS
If you have ever wondered about the art and science of map making (cartography) or the interactions of people and environments, Intro to GIS may be the class for you! Learn the theories behind and practical applications of Geographic Information Systems (GIS) in GEOG 230 C, Intro to GIS. This course provides a broad overview of GIS applications, emphasizing the real-world skills that are in high demand in the private and public sectors.
- ArcGIS.com
Web GIS from ESRI. Sign up for a free account to get a feel for GIS.
- GIS Stack Exchange
GIS stack exchange for GIS professionals and students alike.
- Cal-Atlas
The California Geoportal provides easy and convenient access to freely distributed spatial data for California.
Free spatial data at the national and subnational level for the entire world.


text:
I just saved the U.S. economy, and it only took a minute and a half! Yay me.
Well, I didn't actually save it, but after playing a quick round of the free casual game "Trillion Dollar Bailout!" an image of a newspaper appeared on my screen with the headline "Economy Saved! World Rejoices!!!" Apparently, amid all the disagreement over how the $787 billion economic stimulus package should be divvied up, I managed to make wise choices as to who and what should get funding. A regular Paul Krugman I am.
Ah, if only rescuing the faltering economy were as easy as this newly posted little title on AddictingGames makes it out to be. You just decide whether you want to deliver a bag of money or a slap in the face to the CEOs and homeowners who pop up along New York's skyline and city streets holding signs asking for cash.
It's pretty clear what sort of choices will net you a win. "Punish greedy fat cats and save honest peoples! Hand out moneys to homeowners. Put the hurt on dudes in suits! Do it right and save the world!" reads the game's introduction screen (and no, that wasn't me who put an "s" at the end of people and money).
Sadly, playing this game won't save homeowners from foreclosure or bail out the banks, but it's a harmless enough way to pass a few minutes.
AddictingGames, by the way, also recently brought us the free online game "Hero on The Hudson," which tasks players with emulating the success of US Airways pilot Chesley Sullenberger landing a passenger jet on water.


text:
Course Title: First Aid at Work
Course Duration: 3 Days (18 hours contact time)
Fee per Person: £220
Remote Learning: Not Available
It is a requirement of the Health & Safety law that employers provide a safe and healthy environment to work in. If your assessment of first aid needs highlights the need for qualified first aiders, then this qualification will provide you with suitable, highly trained personnel.
This comprehensive three-day course covers a wide range of first aid emergencies, enabling all learners to deal with emergency situations with confidence in a prompt, safe and effective way. In addition, this course will include any protocol changes that may have arisen since their last training course.
This course will give the learners a qualification to the highest level of First Aid, meeting the statutory requirements of the Health and Safety (First Aid) Regulations 1981.
A range of subjects are covered including –
- Legalities, responsibilities and reporting
- Assessment of the situation
- Dealing with an unresponsive casualty
- Resuscitation
- Use of an AED
- Choking
- Control of bleeding
- Bandaging
- Shock (including Anaphylaxis)
- Fractures and spinal injuries
- Sprains and strains
- Burns
- Heart attacks
- Head injuries
- Stroke
- Chest injuries
- Poisoning
- Eye injuries
- Asthma
- Epilepsy
- Diabetes
Summative practical assessment is ongoing by the instructor each day, along with a written assessment on day 3.
A three year Award in First Aid at Work will be issued to the learner, subject to successful assessment.
A maximum of 12 learners are allowed, they must be a minimum of 14 years of age and a certificate can be offered to all, subject to assessment.
A learner cannot assume a responsibility in the workplace until they reach the age of 16, and then it is the employer's responsibility to ensure that the student is suitable for that role.
The certificate will expire as dated on the certificate, irrespective of age.


text:
Year 5 Class
Learning Letters
Key Information
In Year 5, we are taught by Mrs Entwistle and we are supported by Miss Lincoln and Mr Ashworth (mornings only). Mrs Room also teaches us when Mrs Entwistle is completing her planning and preparation.
We follow the school ethos 'Christ at the Heart of All We Do' to ensure that we can complete our learning to the best of our ability.
I am glad to be returning to Year 5 after an enjoyable maternity leave and have had a lovely warm welcome from the children already. Miss Lincoln has joined Year 5 this year to support us, along with Mr Ashworth in the mornings and Mrs Connolly is with us in the afternoons. Mrs Room continues to take Year 5 on a Monday afternoon for PE and PHSE.
Let me remind you of some key information that will ensure we finish a successful year together...
On Mondays, spellings will be sent home in yellow books; the children will need to learn these for a test on the Friday of the same week. I have demonstrated a range of strategies to learn them, and I shall collect results as a means of assessing.
Additional pieces of work will be sent home, usually on a Friday, that relate to topic or Maths or English, and will generally be due the following Friday. Please see homework as important: it will always be intended to build on the class learning or to prepare for a sequence of work, so support you child in completing it to the best of their ability. That said, please do not over-stress the homework - if, after half an hour, it is proving too much for any of you, then leave it and send in a note. I'll understand. I like to see that children have had a good go, not necessarily a perfect piece at the end. After all, there is usually lots of other jobs to be getting on with at home.
Please listen to your child read at home, and sign and comment in their reading record. This will help us monitor content and progression through the schemes and allows a channel of communication. Children should be taking their assigned 'Home Reading' book home with them every night and returning with it the following day. We are looking for at least 2 signatures in a week. In addition, we will hear your child read each week during 'guided reading' sessions, as well as set and discuss follow-up tasks, free reading and book reviews.
Other activities
PE will take place on Monday pm and on Thursday am. Please go to every effort to ensure children have a full regulation kit on those days. If you have any problems, please call in at the office - I'm sure we can help.
In the meantime, please don't hesitate to get in touch if you have any questions or concerns.
Thank you for your co-operation and we hope you enjoy checking back into the Year 5 pages!
Aliens are landing, or are they? Year 5 are going on a journey through time and outter space. How will they survive? Watch this space.....


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What do glacial surfaces and tea have in common? The obvious answer to a glaciologist… 'a glaciologist – standing on the surface of a glacier, drinking tea'. While this is, of course, correct, here's another that is more interesting and of high relevance to the Dark Snow Project; the surface's colour. Or – more precisely – its pigmentation.
The Dark Snow Project is interested in all things dark on the surface of glaciers and ice sheets. Joe Cook summarizes the ways microbes can contribute to the darkening of glacial surfaces. I'll focus on a specific process that may be the key biotic factor for albedo reduction of the Greenland ice sheet.
There are three species of algae that grow on glacier surfaces worldwide – Cylindrocystis brébissonii, Mesotaenium berggrenii, and Ancylonema nordenskiöldii. I'll call them 'ice algae'. Each belongs in a single group of green algae called Zygnematophyceae, also called conjugating algae due to their inventive and esthetic way of having sex. It is still a mystery why only these few species reside on ice, and from only one group, but we begin to understand how: it's to do with tea!
Imagine the environment of the surface of the Greenland ice sheet… It's of course cold, around freezing all the time, and it can be very bright, further amplified by ice crystals reflecting the incoming light in all directions and multiple bounces between clouds and the surface increases the glare. If you're an alga, then you too are cold and you're dazzled much of the time. You get most of your food from the atmosphere as carbon dioxide, but you still need some other nutrients such as nitrogen and phosphorus, and there is not much of that around. And you may even need to protect yourself from large predators that stalk these vast white planes. Ok, not large for humans. Microscopical predators. But still scary.
It seems that ice algae have solved these problems in one fell swoop by producing a special pigment stored small vesicles (called vacuoles) inside the cells, and its colour has been described (see Yallop et al. 2012). Now, depending on the observers' state of colour-blindness and/or gender, the pigment is seen as purple brown (Yallop et al. 2012), brownish (Remias et al. 2012), and dark brown (Uetake et al. 2010). I am going to call it 'dark'. Until recently, the chemical nature of this dark pigment was unknown. But then Daniel Remias and his colleagues from Innsbruck University decided to look at one of the ice algae – M. berggrenii from an Alpine glacier – more closely (Remias et al. 2012). They managed to resolve the structure of the main compound responsible for the dark colour and gave it a beautiful name: purpurogallin carboxylic acid-6-O-b-D-glucopyranoside.
What can purpurogallin carboxylic acid-6-O-b-D-glucopyranoside do? Well, it's dark, so of course it absorbs sunlight, mostly in the ultraviolet and visible parts of the spectrum. In other words, it is a sunscreen, a protection against the harmful UV radiation and also excessive visible radiation which can inhibit photosynthesis in the cells. But that's not all. The pigment may also represent a sink for surplus energy that cannot be invested in cells due to limitations in temperature or nutrient availability, and may even act as a chemical defense against grazers as, for example, phenolic compounds in marine kelp. So, given the nuisances you have to put up with as an alga living on the surface of an ice sheet, it seems like a very useful thing to have.
And strangely, purpurogallin carboxylic acid-6-O-b-D-glucopyranoside is a pigment that has only been detected in higher plants, such as fermented plant tissue… leaves, more specifically… fermented leaves of Camellia sinensis, even more specifically. Also known as black tea.
So here we are, glaciologists standing on the melting surface of the Greenland ice sheet, sipping tea that is black precisely for the same reason why the ice surface is getting dark – a simple pigment produced by a living organism.
We promise we won't spill much of the tea.
Uetake J, Naganuma T, Hebsgaard MB, Kanda H, Kohshima S (2010) Communities of algae and cyanobacteria on glaciers in west Greenland. Polar Science 4: 71–80
Remias D, Schwaiger S, Aigner S, Leya T, Stuppner H, Lütz C (2012) Characterization of an UV- and VIS-absorbing, purpurogallin-derived secondary pigment new to algae and highly abundant in Mesotaenium berggrenii (Zygnematophyceae, Chlorophyta), an extremophyte living on glaciers. FEMS Microbiology Ecology 79: 638–648
Yallop ML, Anesio AM, Perkins RG, Cook J, Telling J, Fagan D, MacFarlane J, Stibal M, Barker G, Bellas C, Hodson A, Tranter M, Wadham J, Roberts N (2012) Photophysiology and albedo-changing potential of the ice algae community on the surface of Greenland Ice Sheet. ISME Journal 6: 2302–2313


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Heat pumps are machines that move heat from a cool place to a warm place. For example, in the winter, a heat pump will move the heat in the colder outsides into the warm interior living space. Similarly, during hotter months when the house is kept cool, a heat pump will remove the heat from the inside of the house and release it into much warmer outside environment. They do not generate heat, unlike a gas or electric furnace; they simply move it. Heat pump repair is as easy as calling an HVAC specialist.
Traditional heat pumps (where the pump is the only source of heat) are only right in certain situations. Those in colder climates will not see much benefit from heat pumps, as the pump loses effectiveness as the outside temperature drops. In a milder climate, where "cold" means 50° or 40°F, the outside temperature remains high enough to be useful, but at temperatures under 30°F, an additional heat source is needed.
Hybrid Heat Systems
Hybrid heat systems use a heat pump and a furnace to control interior temperature. When the temperature is above 30°F, the pump does the work. When the temperature falls below that point, the furnace takes over. This type of setup is perfectly suitable for colder climates and is much more efficient and effective than just having one or the other.
Heat pumps are rated for efficiency by their HSPF, heating seasonal performance factor. A higher number means less energy consumption and/or waste. There are three different performance options when buying a heat pump—single-stage, two-stage, and modulating heat. Most businesses offer at least two of three types in their product lines, with an example being Goodman's electric heat pump.
Single-Stage Heat Pump
These heat pumps run at full capacity all the time. There is no option to moderate or increase the amount of heat entering the home; when the ideal temperature is reached, the compressor stops and the fan blows out any remaining hot or cold air. Single-stage heat pumps are the most inexpensive option, but they lack the efficiency of the two-stage and modulating varieties.
Two-Stage Heat Pump
A two-stage heat pump has the ability to run at two different levels—maximum capacity and 65% of the maximum capacity. Most homes can maintain a comfortable temperature by keeping the pump on its low setting. Two-stage pumps fall between single stage and modulating pumps in terms of both price and efficiency.
Modulating Heat Pump
Modulating heat pumps are the latest in heat pump technology and, as such, are the highest in price and efficiency out of the three. These types of pumps can run at a range of different capacities, usually from about 40% to 100% are typically best choice for those who want to maintain an ideal living environment all year and save some money whilst doing so.
Heat pumps use refrigerants to facilitate the transport of heat from one location to another. Up until recently, the standard chemical was R22, or freon. However, because it is quite harmful to the environment, most brands have ceased using it and adopted more environmentally-friendly methods. It is set to be completely phased out by 2020; until then, freon is expected to become more and more scarce, as companies stop purchasing it and use up their existing stocks.
Gulf Coast Heating and Cooling is your Heat Pump Specialist on the Mississippi Gulf Coast.


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Available rooms from Thu 29 October 2020 to Fri 30 October 2020.
This hotel is situated in the centre of The Hague only 150 metres from the Bierkade Tram Stop. Mercure features a 24-hour front desk room service and car rental service.
All of the sound-proofed rooms at Mercure Hotel Den Haag Central offer cable TV a refrigerator and a work desk.Guests can fill their refrigerator with a selection of drinks and snacks from the M-Shop kiosk in the reception.
The restaurant M'Biance at Mercure Hotel Den Haag Central serves International light meals and snacks in a modern setting. Guests can enjoy cocktails and a selection of wines from the "flavours from the cellar" menu in casual surroundings at the bar.
The Mauritshuis Museum is less than a 10-minute walk from the hotel. Panorama Mesdag is 20 minutes' walk away. The seaside town of Scheveningen is 4.5 km away and reachable by tram.
- Restaurant
- Laundry
- Meeting/Banquet Facilities
- Newspapers
- Parking
- Room Service
- Safety Deposit Box
- Air Conditioning
- Bar
- Currency Exchange
- 24-Hour Front Desk
- Express Check-In/Check-Out
- Dry cleaning
- Internet services
- Soundproof rooms
- Non-smoking throughout
- Breakfast in the room
- Heating
- Fax/photocopying
- Luggage storage
- WiFi
- Restaurant (à la carte)
- Private check-in/check-out
- golf course (within 3 km)
- car hire
- Facilities for Disabled Guests
- Non-smoking Rooms
- WiFi available in all areas
- Business centre
- Free WiFi
- Lift


text:
Acoustic Imaging of Perforation Erosion in Hydraulically Fractured Wells for Optimizing Cluster Efficiency
- Authors
- Stephen Robinson (DarkVision Technologies Inc.) | Thomas Littleford (DarkVision Technologies Inc.) | Tim Luu (DarkVision Technologies Inc.) | Kacper Wardynski (DarkVision Technologies Inc.) | Andrew Evans (Jagged Peak Energy) | Blake Horton (Encana Oil & Gas, USA) | Michael Oman (Petronas Canada)
- https://doi.org/10.2118/199718-MS
- Document ID
- Publisher
- Society of Petroleum Engineers
- Source
- SPE Hydraulic Fracturing Technology Conference and Exhibition, 4-6 February, The Woodlands, Texas, USA
- Publication Date
- 2020
- Document Type
- Conference Paper
- Language
- English
- 978-1-61399-698-0
- Copyright
- 2020. Society of Petroleum Engineers
- Keywords
- Perforation Measurement, Wellbore Imaging, Cluster Efficiency, Perforation Erosion, Acoustic Imaging
- Downloads
- 42 in the last 30 days
- 1,168 since 2007
- Show more detail
- View rights & permissions
|SPE Member Price:
|SPE Non-Member Price:
A new solid-state, high-resolution acoustic imaging technology has been applied in hydraulically fractured wells to image and quantify perforation erosion. The downhole device captures detailed full-lateral logs of horizontal wells, without the need for clear fluids to facilitate measurements. This paper discusses how the imaging technology functions, lab testing that validated the measurements, and field testing completed with several operators in a variety of reservoirs and basins.
This high-resolution, acoustic-based downhole imaging technology provided a 360-degree view inside a variety of wells. Instead of optics, the imaging technology used high-frequency sound waves to image a full lateral through opaque fluids at sub-millimetric levels. The imaging tool was conveyed on either tractor or e-coil to continuously log the wellbore between 15 and 30ft/min, completing a full azimuthal scan that imaged individual perforations regardless of orientation. A range of imaging techniques quantified and measured each perforation to determine the extent of erosion in each perforation, cluster, and stage sustained during the hydraulic fracturing process.
The acoustic imaging technology was initially tested in the lab with calibration jigs and perforated pipe samples to validate its accuracy. The technology was then field deployed by several operators and used to assess perforation erosion in extended horizontal wells by scanning the entire well from toe to heel and measuring thousands of perforations in a single log. The data gathered quantified the perforation diameter, perforation erosion range, perforation orientation dependency, cluster efficiency, perforation erosion bias, and discovered the presence of significant casing damage around areas where plugs were set. The image results showed the distribution, size, and measured diameter for each perforation in each cluster. The results noted toe or heel biases and how these changed in different well types and designs in different basins. The data allowed operators to reach conclusions regarding erosion and cluster efficiency across the entire lateral in various wells in the Montney, Permian, and Anadarko basins. Wells across North America were scanned in several other basins for comparison purposes using various completion designs from different operators.
The high-resolution acoustic imaging technology offers a robust 360-degree view of long horizontal wells and perforation measurements post-fracture. Over 35,000 perforations have been measured using the technology in the field across various well types with different designs. The technology has proven to be a valuable tool in improving and optimizing completion designs by providing detailed feedback on where designs are working effectively and where they can be improved.
|File Size
|Number of Pages
Cramer, D.D. 1987. The Application of Limited-Entry Techniques in Massive Hydraulic Fracturing Treatments. SPE 16189-MS presented at the SPE Production Operations Symposium, Oklahoma City, 8-10 March. http://dx.doi.org/10.2118/16189-MS
Cramer, D.D., Friehauf, K., Roberts, G.. 2019. Integrating DAS, Treatment Pressure Analysis and Video-Based Perforation Imaging to Evaluate Limited Entry Treatment Effectiveness. SPE-194334-MS presented at the SPE Hydraulic Fracturing Technology Conference and Exhibition, The Woodlands, Texas, USA, 5-7 February 2019. https://doi.org/10.2118/194334-MS
Crump, J.B., Conway, M.W. 1988. Effects of Perforation-Entry Friction on Bottomhole Treating Analysis. J Pet Technol 40 (08). 1,041 - 1,048. SPE-15474-PA. http://dx.doi.org/10.2118/15474-PA
Roberts, G., Lilly, T. B., & Tymons, T. R. 2018. Improved Well Stimulation Through the Application of Downhole Video Analytics. SPE-189851-MS presented at SPE Hydraulic Fracturing Technology Conference and Exhibition, 23-25 January 2018, The Woodlands, Texas, USA http://dx.doi.org/10.2118/189851-MS
Wu, C-H., Sharma, M.M. 2016. Effect of Perforation Geometry and Orientation on Proppant Placement in Perforation Clusters in a Horizontal Well. Presented at the SPE Hydraulic Fracturing Technology Conference, The Woodlands, Texas, USA, 9-11 February 2016. SPE-179117-MS. http://dx.doi.org/10.2118/179117-MS
Yi, S., Wu, C-H., Sharma, M. 2019. Optimization of Plug-and-Perf Completions for Balanced Treatment Distribution and Improved Reservoir Contact. Presented at the SPE Hydraulic Fracturing Technology Conference, The Woodlands, Texas, USA, 5-7 February 2019. SPE-194360-MS. https://doi.org/10.2118/194360-MS


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A good massage therapists in London.
I don't know about you how you love your getaways, but I will tell you my secret. Visiting the best massage therapist. See, when you go to a massage therapist, you know what you want. Your body will appreciate the tender care and love you will give. You need a therapist that is sensual with the tender parts of your body in an experienced manner.
You basically need a show of talent in your services. I tell you what, a massage therapist London knows exactly how to deliver your needs. Like I said, I love my hangouts with massage therapists, a good one for that matter. And to have coming back to your spar, again and again, you have to earn it. You have to show the talent in your skill. Show me the passion you have earned from your experience as a massage therapist.
This is the exact kind of service you will get when you visit a massage therapist in London. Visiting tantric temple for the first time, you will be awed by the pleasant reception you will get. The way they communicate with you as a customer will have you feeling at home already. The fact that they understand that good communication skills is important to make the clients day even better will get you hooked.
This makes them the best massage therapist London of all London has to offer. Another thing is the skill used by the therapists. It is like they understand the needs of your body better than you understand yourself. All you have to do is sit there and relax and wait for your body to feel like alive all over again. Tantric temple in London has high skilled therapists that understand how to incorporate different techniques and approaches to the body of a client. With them, satisfaction is guaranteed. Now, let me tell you about something you may not know.
You don't know what an experienced massage therapist works until you have visited Tantric Temple. Each and everyone one of them provide a service that qualifies them to be the best massage therapist in London. The perfection of their work is shown in the feeling of your body when you leave, because I tell you, you will be begging to stay. Tantric temple provides the best massage therapists in London known for their emphatic and sensual working.
Being empathic allows them to understand and be receptive to your feelings and wants as a client. The constant assurance and being able to meet your therapy needs for every massage therapy session. And the sensual aspect in which they execute their services to their clients is just fulfilling. It makes you understand your body better as you continue the exercise.
Let me not go on, but I will leave you with this; if you are you ever London or live in London, and you want that body treatment you have been longing for forever, try visiting the best massage therapist London. I promise you, you will fall in love with the Tantric Temple.


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Environmentally Conscious Wineries Choose Verallia's ECO SeriesTM Wine Bottles
Since introducing the first eco-conceived wine bottle in North America just two short years ago, Verallia has sold over 180 million ECO SeriesTM bottles. Although the primary intent of the ECO SeriesTM bottle was to provide customers with a high quality bottle with less overall impact on the environment, over 300 environmentally conscious wineries are capitalizing on the sustainability characteristics of the bottle and widely-accepted marketing appeal the bottle has with consumers.
"We have had an overwhelming response to the ECO SeriesTM from our wine customers, both large and small," said Bob Parise, Vice President, Sales and Marketing, Wine Sector for Verallia North America. "In 2009, we introduced the bottle as part of a company-wide sustainability initiative. Although we knew the wine industry already had a deep commitment to the environment, we learned very quickly the industry has a keen ability to market that commitment to environmentally conscious consumers."
Finding great success in marketing wine in the ECO Series™ bottle, Eight Arms Cellars in Suisun Valley, Calif. is set to bottle all of its wines in Verallia's ECO SeriesTM bottles in 2011. Eight Arms is a small winery known for its Syrah wines and lives by its brand "Go Green, Drink Red". After using the ECO SeriesTM Evolution bottle for packaging "The Tentacle" wine last spring, Eight Arms experienced an overwhelming response from consumers selling out of the product in less than 60 days.
"I first chose the ECO SeriesTM bottle for The Tentacle Syrah last spring because the bottles are more carbon friendly and made locally with recycled glass," said Boltin.
On a much larger scale, Goose Ridge Estate Vineyard and Winery is located in the heart of the Washington's Columbia Valley. The third-generation, family-owned and operated winery practices sound viticulture principles and manages every aspect of winemaking from planting to harvesting and processing to packaging. As a result of the family's commitment to excellence, the winery's 2006 Vireo was ranked #41 in the Wine Spectator's Top 100 of 2010.
"We take pride in every bottle of wine we produce, and want to ensure every bottle is one that we'd proudly serve our own family and closest friends," said Molly Stutesman, third generation family partner of Goose Ridge Estates. "We chose the ECO SeriesTM for our g3 and Stonecap wines because sustainability is important to our process and our brand."
Named for the third generation of family owners, g3 includes both a premium red wine and a premium white wine blend. Both the g3 and Stonecap wines are 100% estate fruit grown and packaged on premise reducing the winery's carbon footprint.
The ECO SeriesTM allows Goose Ridge to stay competitive in this wine category by offering a local supplier and a high quality, affordable bottle. "We believe the bottle brings added value and benefits to consumers who are looking for producers and products that endorse or use practices that are sustainable and responsible. Using a local supplier makes it that much more sustainable and affordable to bottle our premium wines," added Stutesman.
Verallia's ECO SeriesTM lines were the first glass containers manufactured in North America focusing on eco-conception, a process developed to maintain high quality and enhance customer appeal all the while lessening the overall impact on the environment (energy demand, transportation impact, and CO2 emissions). For example, the ECO SeriesTM Inspiration claret wine bottle, when compared to a "classic" 20 oz claret wine bottle, reduces consumption of energy for manufacturing and CO2 emissions by 19% and further reduces greenhouse gas and impact on transportation by 10% (based on average distance of 200 miles).
"ECO SeriesTM bottles have not only proven to be a success for Verallia but, more importantly, are bringing added value and real success to our customers," said Doug Hesche, Vice President, Market Development for Verallia North America. "It's great to be able to offer customers a product that is not only better for the environment but, also, more affordable and marketable to the end consumer."
The ECO SeriesTM catalog is now available by contacting Kate Gerber at email@example.com
About Verallia and Saint-Gobain Packaging
Verallia is the glass packaging brand of Saint-Gobain created for the glass bottles and jars market. Saint-Gobain Packaging, a major global manufacturer, designs and manufactures glass packaging that is respectful of the environment, is endlessly recyclable, and preserves & promotes the quality of products. Verallia glass is pure and safe, respecting the well-being of consumers.
Verallia North America, the Saint Gobain Packaging Sector's North American business, is a major glass container manufacturer in the United States and produces Verallia brand glass bottles and jars for the wine, beer, beverage, spirits, and food container markets with 13 strategically located plants and a state of the art distribution center.


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The Cambridgeshire Association for Local History is the umbrella organisation for all things local history in Cambridgeshire.
Villagers was awarded one of the Association's Local History Book Awards for 2012 at a 'Celebration of Local and Social History' held at Soham on Saturday 16 June 2012. This impressive framed certificate commemorates the event.
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Although, technological advances in aerial robotics are being made quite rapidly, the opportunity exists for customizing and combining existing technologies to develop cost efficient flying tools that are suitable for the specific needs of our clients. For clients interested in using drones as automated tools for their business operations, Wilstair offers the peace of mind of providing project management assistance to integrate adequate technology that satisfies established requirements. The outcome of this service is to provide our clients a technology demonstrator suitable for their needs. In order to accomplish such task, Wilstair, alongside the client, will source from established drone technology companies the best and most cost efficient products in the market that will satisfy the requirements of the project.
Why is Wilstair willing to support such projects? Simple, because the success of our clients matter to us. In addition to being continuous students of the Unmanned Aerial Systems (UAS) market, we are also looking out for our clients. We want our clients to make educated decisions that captures important factors such as performance, reliability, and safety while remaining cost-centric.


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Monday March 6, 2013 - Day 4
We began by taking the INTA shuttle to the convention center only to realize on arrival that the Opening Ceremony was being held at the Omni Hotel. After brisk walking we made it to the Ballroom for the opening. The Guest speaker Jerry Jones owner of the Dallas Cowboys was both humorous and engaging as he told us how he developed the Cowboy franchise which he bought in 1989 into the second most valuable sports brand in the world. Manchester United is the number 1 brand.
I then attended in the same Ballroom as the opening ceremony the Session on Social Media: Sidestepping Problems and Pitfalls. When that session finished I rushed over to the Exhibit Hall to examine a few booths and then rushed back for the next session Do We Launch: Assessment of the new gTLD Program. After this I found Dianne Daley and we headed for the Millen White, Zelano & Branigan luncheon at the Union Station which was terrific. Great food and award winning beer. We met some new colleagues, were encouraged to go to Hong Kong, participated in a review of the previous days receptions and discussed the upcoming receptions that could not be missed. After this we rushed (a word used frequently at INTA) back for the next session with start time 3:30 pm. My session was Trademark Bullying. I reserve comment on this topic/concept as it appears did the rest of the attendees of the session as when it came time for Q&A no one had any questions.
With the sessions over we headed off to the various receptions. Monday tends to be the most packed day for firm hosted receptions. We began at 5pm by attending the Knobbe Martens reception at the W Hotel, I think on the 33rd Floor. It was a nice reception. We left that reception and went to the Iron Cactus Mexican Grill & Margarita Bar for another reception only to discover that we mixed up the days and the reception is on Tuesday.We were not deterred and simply moved on to the next reception which was held at the Sambuca Restaurant hosted by Honigman. Yet another great reception with wonderful food and drinks. We left that reception went to another and then attended the Haynes Boone reception at the Perot Museum of Nature and Science. Special mention to our friend Phil now the Managing Partner of the firms Washington Office. I have included pictures. Food as usual wonderful catered by ....who else Wolfgang Puck.
At this point I need to bring up the issue of taxis. Dallas has tons of taxis but as I heard one attendee remark they do not appear to know the city. Our cab driver had to use GPS to find the Perot museum!
Our colleague Joanne Wood Rattray and her husband Robin encouraged us to attend the Groth & Co. Reception at the Dallas Aquarium. Excellent reception hosted by A Swedish law firm. For every guest who attended they contributed to a charity in Africa. We saw Manatees, sea snakes, tarantulas, and even penguins...go figure.
We left that reception, and moved to the Arent Fox reception at the Tower Club. Another great view of the Dallas skyline at night. After that we were off to the Merchant & Gould reception. After some dancing we called it quits at 11:30pm.


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Related research
ClimateXChange (CXC)
A collaborative initiative between sixteen of Scotland's leading research and higher education institutions undertaking work on climate change and the transition to a low carbon economy
Climate change risk assessment (CCRA)
This page describes Forest Research's role as Sector Champion for forestry within the UK Government's framework for climate change risk assessment under the Climate Change Act.
This page summarises the work of OneOak, an educational project organised by the Sylva Foundation.
Climate change impacts
Examining the effects of climate change and air pollution on woodland and forestry.
Forestry and climate change mitigation
Research to improve our understanding of how forests and woodlands can contribute to the carbon balance at both local and global scales
'Assessing Resilience' Programme
Work package leader for 'Quantifying Changes to Resilience', which includes much of our measurement and analysis programme on stand-scale carbon and GHG balance. Also working on climate change risk assessment.
'Valuing Ecosystem Services' Programme
Research on including the wider range of impacts that tree planting and woodland creation can have on the climate into assessments.
'Delivering Resilient Forests' Programme
Research and advice on adaptation measures
Coordinating FR input through the Harwood Forest C & GHG balance project.
ClimateXChange project
Coordinator of FR work, and member of project Directorate, linking climate change research and policy needs for Scottish Government and agencies.
Peer reviewed journal articles
Ilie, I., Dittrich, P., Carvalhais, N., Jung, M., Heinemeyer, A., Migliavacca, M., Morison, J.I.L., Sippel, S. , Subke, J-A., Wilkinson, M. & Mahecha, M.D. (2017). Reverse engineering model structures for soil and ecosystem respiration: the potential of gene expression programming. Geoscientific Model Development 10: 3519-3545, doi: 10.5194/gmd-10-3519-2017.
Pinnington, E.M., Casella, E., Dance, S.L., Lawless, A.S., Morison, J.I.L., Nichols, N.K., Wilkinson, M., & Quaife T.L. (2017). Understanding the effect of disturbance from selective felling on the carbon dynamics of a managed woodland by combining observations with model predictions JGR- Biogeosciences, 10.1002/2017JG003760.
Vanguelova, E.I., Pitman, R., Benham, S. Perks, M.,and Morison, J.I.L. (2017). Impact of tree stump harvesting on soil carbon and nutrients and second rotation tree growth in mid-Wales, UK. Open Journal of Forestry7: 58-78. doi: 10.4236/ojf.2017.71005.
Yamulki, S.and Morison, J.I.L. (2017). Annual greenhouse gas fluxes from a temperate deciduous oak forest floor. Forestry 90: 541-552. doi: 10.1093/forestry/cpx008
Wilkinson, M., Eaton, E.L.and Morison, J.I.L. (2017). .European Journal ofForest Research 136: 1-12.doi:10.1007/s10342-016-0998-z
Pinnington, E.M., Casella, E., Dance, S.L., Lawless, A. S., Morison, J.I.L., Nichols, N.K., Wilkinson, M. and Quaife, T.L. (2016). Investigating the role of prior and observation error correlations in improving a model forecast for forest carbon balance using Four-dimensional Variational data assimilation. Agricultural and Forest Meteorology 228: 299-314. doi:10.1016/j.agrformet.2016.07.006.
Wingate, L., Ogée, J., Cremonese, E., Filippa, G., Mizunuma, T., Migliavacca, M., Moisy, C., Wilkinson, M., Moureaux, C., Wohlfahrt, G., Hammerle, A., Hörtnagl, L., Gimeno, C., Porcar-Castell, A., Galvagno, M., Nakaji, T., Morison, J. Kolle, O., Knohl, A., Kutsch, W., Kolari, P., Nikinmaa, E., Ibrom, A., Gielen, B., Eugster, W., Balzarolo, M., Papale, D., Klumpp, K., Köstner, B., Grünwald, T., Joffre, R., Ourcival, J.-M., Hellstrom, M., Lindroth, A., George, C., Longdoz, B., Genty, B., Levula, J., Heinesch, B., Sprintsin, A., Yakir, D., Manise, T., Guyon, D., Ahrends, H., Plaza-Aguilar, A., Guan, J. H. and J. Grace (2015). Interpreting canopy development and physiology using a European phenology camera network at flux sites. Biogeosciences 12: 5995-6015.
Wilkinson, M., Crow, P., Eaton, E.L. and Morison, J.I.L. (2015). Effects of management thinning on CO2 exchange by a plantation oak woodland in south-eastern England. Biogeosciences Discussions 12: 16197-16232. doi: 10.5194/bgd-12-16197-2015.
D'Odorico, P., Gonsamo, A., Gough, C.M., Bohrer, G., Morison, J., Wilkinson, M., Hanson, P.J., Gianelle, D., Fuentes, J.D. and Buchmann, N. (2015). The match and mismatch between photosynthesis and land surface phenology of deciduous forests. Agricultural and Forest Meteorology 214-15: 25-38.
Ward, H.C., Kotthaus, S., Grimmond, C.S.B., Bjorkegren, A., Wilkinson, M., Morrison, W.T.J., Evans, J.G., Morison, J.I.L. and Iamarino, M. (2015). Effects of urban density on carbon dioxide exchanges: observations of dense urban, suburban and woodland areas of southern England. Environmental Pollution 198: 186-200.
Barsoum, N., Eaton, E.L., Levanič, T., Pargade, J., Bonnart, X. and Morison J.I.L. (2015). Climatic drivers of oak growth over the past one hundred years in mixed and monoculture stands in southern England and northern France. European Journal of Forest Research 134: 33-51. doi: 10.1007/s10342-014-0831-5.
Harris, Z., McNamara, N.P., Rowe, R., Dondini, M., Finch, J., Perks, M., Morison, J., Donnison, I., Farrar, K., Sohi, S., Ineson, P., Smith, P. andTaylor, G. (2014). . Biofuels 5: 111-116.
Wang, S., Hastings, A., Wang, S., Sunnenberg, G., Tallis, M. J., Casella, E., Taylor, S., Alexander, P., Cisowska, I., Lovett, A., Taylor, G., Firth, S., Moran, D., Morison, J. and Smith, P. (2014). The potential for bioenergy crops to contribute to meeting GB heat and electricity demands. GCB Bioenergy 6: 136–141. doi: 10.1111/gcbb.12123
Viger, M., Rodriguez-Acosta, M., Rae, A.M., Morison, J.I.L. andTaylor, G. (2013). Toward improved drought tolerance in bioenergy crops: QTL for carbon isotope composition and stomatal conductance in Populus. Food and Energy Security 2: 220-239. Article first published online: 3 DEC 2013, DOI: 10.1002/fes3.39
Vanguelova E.I., Nisbet T.R., Moffat A.J., Broadmeadow S., Sanders, T.G.M.and Morison, J.I.L. (2013). A new evaluation of carbon stocks in British forest soils. Soil Use and Management 29(2): 169-181. doi: 10.1111/sum.12025.
Mizunuma T., Wilkinson M., Eaton, E.L, Mencuccini, M., Morison, J.I.L.and Grace, J. (2013). The relationship between carbon dioxide uptake and canopy colour from two camera systems in a deciduous forest in southern England. Functional Ecology 27: 196-207.,doi: 10.1111/1365-2435.12026.
Tallis M.J., Casella E., Henshall P.A., Aylott M.J., Randle T.J., Morison J.I.L. andTaylor G. (2013). Development and evaluation of ForestGrowth-SRC a process-based model for short rotation coppice yield and spatial supply at a national scale. Global Change Biology Bioenergy 5: 53-66.
Stokes V.J., Morecroft M.D.and Morison J.I.L. (2010). Comparison of leaf water use efficiency of oak and sycamore in the canopy over two growing seasons. Trees – Structure and Function 24: 297-306.
Book and book chapters (including published conference proceedings)
Grace, J., Morison, J. & Perks, M. (2013) Forests, Forestry and Climate Change, Chap 10 in Challenges and Opportunities for the World's Forests in the 21st Century, T. Fenning, (Ed). Forest Sciences, Vol. 81, Springer., DOI 10.1007/978-94-007-7076-8_1
FC and FR publications including official statistics
Vaz Monteiro, M., Handley, P. Morison, J.I.L., Doick, K.J. (2019). The role of urban trees and greenspaces in reducing urban air temperatures. Forest Research Research Note, FCRN037, 12pp.
Morison, J., Matthews, R., Miller, G., Perks, M., Randle, T., Vanguelova, E., White, M. and Yamulki, S. (2012). Understanding the carbon and greenhouse gas balance of forests in Britain (PDF-7422K). Forestry Commission Research Report 18. Forestry Commission, Edinburgh. i–vi + 1–149 pp.
Vanguelova, E., Broadmeadow, S., Anderson, R., Yamulki, S., Randle, T., Nisbet, T. & Morison, J. (2012) A Strategic Assessment of the Afforested Peat Resource in Wales. Report for Forestry Commission Wales, Project No. 480.CY.00075, October 2012., 141 pp.
Morison, J.I.L., Vanguelova, E., Broadmeadow, S., Perks, M., Yamulki, S.and Randle, T. (2010). Understanding the GHG implications of forestry on peat soils in Scotland (PDF-2140K). Report for Forestry Commission Scotland, October 2010, Forest Research, 56pp.
Forestry Commission (2011). Forests and Climate Change. UK Forestry Standard Guidelines. Forestry Commission, Edinburgh. i–iv + 1–68 pp. ISBN 978-0-85538-832-4. (member of drafting panel).
Ray, D. Morison, J.and Broadmeadow, M. (2010). Climate change: impacts and adaptation in England's woodlands (PDF-1920K) . Forestry Commission (England), Research Note FCRN 201, 16pp. Forestry Commission, Edinburgh.
Contract and project reports –excluding any which are commercially confidential
Perks, M. Khomik, M., Bathgate, S., Chapman, S., Slee, B., Yeluripati, J., Roberts, D. & Morison, J. (2018). Agroforestry in Scotland – potential benefits in a changing climate. Report for ClimateXChange, Edinburgh, 48pp.
Morison, J. I. L. and Matthews, R. B. (eds.) (2016): . 24pp, ISBN 978-0-9934074-0-6, and associated technical papers,
Climate Change Risk Assessment Evidence Report, 2016. Contributing author for Chapter 3, "Natural Environment and Natural Assets", lead author Iain Brown. Published July 2016, c. 200pp.
ECI, HRW, Climate Resilience and Forest Research (2013) Assessing preparedness of England's natural resources for a changing climate: exploring trends in vulnerability to climate change using indicators. Final Report to the Adaptation Sub-Committee, June 2013. (contribution to forestry section).
Morison, J.I.L. (2012) Afforested Peatland Restoration. "Enquiry Response" for ClimateXChange, Scotland. Edinburgh, 8pp.
Moffat, A.J., Morison, J.I.L., Nicoll, B. and Bain, V. (2012). Climate Change Risk Assessment for the Forestry Sector. Defra, London. 182pp.
Trade journal and newsletter articles
Vanguelova, E., Morison J., Nisbet, T. & Martin, S. (2015) Taking stock of forest soil carbon. Forestry & Timber News, August 2015.
Vanguelova, E., Moffat A.., Morison J. & Martin, S. (2011) Can we manage the impacts of changing climate on our forest soils ? Forestry & Timber News, June 2011.
Hemery, G. & Morison J. (2011). The science behind the OneOak project. Chartered Forester. Winter 2010, p. 12-14.
Scientific conferences: presentations or posters
Freer-Smith, P. & Morison, J.I.L. (2016) Climate change and the sustainable intensification of planted forests. Presentation to:; EFI Atlantic/IUFRO, IEFC, Biarritz, 13th June 2016.
Morison, J.I.L., Moffat A.J. & Gazzard R. (2014) Addressing the risks from climate change, particularly wildfire. Presentation to: Improving forest resilience, ICF SE Regional Conference, 27th November 2014, Windsor.
Grace J, & Morison, J.I.L. (2012) Sitka spruce as a sink for carbon dioxide. Presentation to: Managing forests for ecosystem services: can spruce forests show the way? IUFRO, Edinburgh, October 8th -11th 2012.
Morison, J.I.L. (2012) The possible contribution of trees, woods and forests to the UK GHG balance. Presentation to: CARBiFOR II Stakeholder meeting, 4th September, Dublin.


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This is not my first post about the many reasons why I love faux-liage but since then I now have twice as many faux plants in my house which has been entirely fuelled by their popularity on the high street. Whereas it was once far from ok to have fake flowers in your home, now it's positively welcomed and with good reason. For those of us that weren't blessed with green fingers we can still enjoy a home filled with greenery that actually looks healthy and requires no watering.
One trick I've learned along the way is to choose foliage over flowers when it comes to fakery. Unfortunately I find flowers (that are affordable) are not as convincing as plants. On the other hand succulents and cacti look amazing and perhaps it has something to do with the fact that the real ones look surprisingly fake in the first place! If you saw my recent post about Matalan, (you can read it here) you would have seen the little faux aloe plant I bought – it resides next to real plants and no one has ever questioned it!
I've also come to realise faux-liage is great in places where there's no chance a real plant would survive. Our downstairs toilet for instance has a window in it the size of a cat flap, and since I painted it navy any natural light that does make it through has to fight to be noticed. My first fake purchase, the cheese plant leaves from Ikea, now reside and thrive in there and look quite wonderful against the blue.
The final piece of wisdom I would impart about faux-liage is to put them in vases that don't show off the stems. I love my bunch of eucalyptus dearly but sadly I do not have the same love for the clumpy plastic stems that I had to contort to fit into this vase they stand in. The cheese plant stems are not wholly unattractive so they are sitting in a glass vase for now but I do have a ceramic vase waiting for them when I get the chance to swap them. Also be warned that fake soil is not as far advanced as the greenery yet so don't be afraid to put your plant in a bigger pot or cover the base with stones to disguise the plastic soil.
Obviously I still have a lot of love for a bunch of blooms and this time of year especially when I can fill my house with daffodils and tulips every time I visit the supermarket, but my fondness for fakery is strong – and for good reason – they are so good these days and affordable too! I've put together my edit of the best plants that are available now.
Happy Shopping
This post was not sponsored, all thoughts and opinions are my own.