Bacterial Physiological State in Wastewater: Monitoring Maintenance and Production with Leu/TdR Ratio for Less Pollution

In domestic wastewater, bacterial physiology controls cell production (growth, replication) and cell maintenance, determining how energy is allocated between these two processes. The aim here was to develop a method to quantify these cellular processes so that the bacterial physiological state could be manipulated to lower this source of pollution. We simultaneously used the incorporation of radiolabelled thymidine into DNA (a measure of new cell synthesis) and leucine into protein in wastewater to quantitatively distinguish bacterial growth from maintenance processes. We found that DNA and protein syntheses were coupled in wastewater after substrate enrichment (with glucose or acetate)??balanced growth. Once the substrate was depleted, the two processes became uncoupled??unbalanced growth. In this physiological state, the bacteria were synthesising protein, but fewer bacteria were replicating. More energy was allocated to cell maintenance than replication. A mean Leu/TdR ratio of 7.4 was determined for wastewater and was similar to natural aquatic ecosystems. As the bacterial growth rate decreased, the Leu/TdR ratios increased. We show how the simultaneous measurement of [³H]Leu and [³H]TdR quantitatively distinguishes balanced from unbalanced growth. Low [³H]Leu/[³H]TdR ratios indicated bacteria were physiologically stressed, an ideal state for biological wastewater treatment processes (WWTP) as the bacteria divert more energy to maintenance activities instead of growth. Leu/TdR ratios of 70 have been recorded in natural aquatic ecosystems which suggests WWTP have potential to be manipulated to achieve much higher Leu/TdR ratios than we report here. Changes to plant operation to improve operation efficiency include finding the optimum rate of substrate (pollution supply) or alternating aerobic and anaerobic periods to maximise the Leu/TdR ratio to achieve less biomass production for land disposal and more cost-effective operation that generates less pollution in the effluent.     

Collagen fibril alignment and deformation during tensile strain of leather: a small-angle X-ray scattering study

The distribution and effect of applied strain on the collagen fibrils that make up leather may have an important bearing on the ultimate strength and other physical properties of the material. While sections of ovine and bovine leather were being subjected to tensile strain up to rupture, synchrotron-based small-angle X-ray scattering (SAXS) spectra were recorded edge-on to the leather at points from the corium to the grain. Measurements of both fibril orientation and collagen d spacing showed that, initially, the fibers reorient under strain, becoming more aligned. As the strain increases (5-10% strain), further fibril reorientation diminishes until, at 37% strain, the d spacing increases by up to 0.56%, indicating that significant tensile forces are being transmitted to individual fibrils. These changes, however, are not uniform through the cross-section of leather and differ between leathers of different strengths. The stresses are taken up more evenly through the leather cross-section in stronger leathers in comparison to weaker leathers, where stresses tended to be concentrated during strain. These observations contribute to our understanding of the internal strains and structural changes that take place in leather under stress.     

Pesticide extraction efficiency of two solid phase disk types after shipping

An interlaboratory study was conducted to compare pesticide recovery from Empore C(18) and Speedisks C(18)XF solid phase extraction disks after shipping. Four pesticides were used for the comparison of the two disk extraction materials: atrazine, diazinon, metolachlor, and tebuconazole. These pesticides were chosen to provide a range of physiochemical properties. Water samples were extracted onto the disk types and shipped to a cooperating laboratory for elution and analysis. The mean recoveries from Empore disks were atrazine, 95%; diazinon, 91%; metolachlor, 92%; and tebuconazole, 83%. The recoveries from Speedisks C(18)XF were atrazine, 89%; diazinon, 87%; metolachlor, 86%; and tebuconazole, 79%. Means for each of the pesticides using the different disk types were not statistically different (alpha = 0.05), but results were more variable when using Speedisks C(18)XF as compared to Empore disks. Reasons for the increased variability are discussed, but overall results indicate that Speedisks C(18)XF could be used as an alternative to Empore disks. Speedisks C(18)XF are enclosed in a plastic housing, so they can be used more easily in remote sampling sites without the possibility of glassware breakage, no prefiltration of samples is needed, and there are realignment problems that can be associated with the Empore disks.     

Human-carnivore conflict in China: a review of current approaches with recommendations for improved management

Human-wildlife conflict (HWC) is a conservation concern that increasingly threatens the continued existence of some of the world's most endangered species. With an increase in human population, urban sprawl and subsequent encroachment on wild land, human and wildlife interaction has become inevitable. In the majority of cases, this interaction results in a negative outcome for humans, wildlife or both. In China, these key elements, along with a decrease in wild prey species, have resulted in the expansion of HWC encounters, and the need for alleviating this conflict has become a conservation priority. Loss of human life, livestock and/or crops is most often the catalysts that fuel HWC. Techniques to alleviate conflict around the world have included preventative measures and mitigation techniques, such as financial compensation and other incentive programs. Both types of measures have had variable success. We review the current status of human-carnivore conflict management in China, and, drawing lessons from around the globe, we make recommendations for improving conservation management in China. For example, an increase in law enforcement in nature reserves is vital to reducing human disturbance in prime carnivore habitat, thereby reducing conflict encounters. Also, modifications to current wildlife compensation programs, so that they are linked with preventative measures, will ensure that moral hazards are avoided. Furthermore, investigating the potential for a community self-financed insurance scheme to fund compensation and increasing efforts to restore wild prey populations will improve the outcome for wildlife conservation. Ultimately, HWC management in China will greatly benefit from an integrative approach.     

Identification of stable normalization genes for quantitative real-time PCR in porcine articular cartilage

Background

Expression levels for genes of interest must be normalized with an appropriate reference, or housekeeping gene, to make accurate comparisons of quantitative real-time PCR results. The purpose of this study was to identify the most stable housekeeping genes in porcine articular cartilage subjected to a mechanical injury from a panel of 10 candidate genes.

Results

Ten candidate housekeeping genes were evaluated in three different treatment groups of mechanically impacted porcine articular cartilage. The genes evaluated were: beta actin, beta-2-microglobulin, glyceraldehyde-3-phosphate dehydrogenase, hydroxymethylbilane synthase, hypoxanthine phosphoribosyl transferase, peptidylprolyl isomerase A (cyclophilin A), ribosomal protein L4, succinate dehydrogenase flavoprotein subunit A, TATA box binding protein, and tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein—zeta polypeptide. The stability of the genes was measured using geNorm, BestKeeper, and NormFinder software. The four most stable genes measured via geNorm were (most to least stable) succinate dehydrogenase flavoprotein, subunit A, peptidylprolyl isomerase A, glyceraldehyde-3-phosphate dehydrogenase, beta actin; the four most stable genes measured via BestKeeper were glyceraldehyde-3-phosphate dehydrogenase, peptidylprolyl isomerase A, beta actin, succinate dehydrogenase flavoprotein, subunit A; and the four most stable genes measured via NormFinder were peptidylprolyl isomerase A, succinate dehydrogenase flavoprotein, subunit A, glyceraldehyde-3-phosphate dehydrogenase, beta actin.

Conclusions

BestKeeper, geNorm, and NormFinder all generated similar results for the most stable genes in porcine articular cartilage. The use of these appropriate reference genes will facilitate accurate gene expression studies of porcine articular cartilage and suggest appropriate housekeeping genes for articular cartilage studies in other species.     

Trypanocidal Activity of Marine Natural Products

Marine natural products are a diverse, unique collection of compounds with immense therapeutic potential. This has resulted in these molecules being evaluated for a number of different disease indications including the neglected protozoan diseases, human African trypanosomiasis and Chagas disease, for which very few drugs are currently available. This article will review the marine natural products for which activity against the kinetoplastid parasites; Trypanosoma brucei brucei, T.b. rhodesiense and T. cruzi has been reported. As it is important to know the selectivity of a compound when evaluating its trypanocidal activity, this article will only cover molecules which have simultaneously been tested for cytotoxicity against a mammalian cell line. Compounds have been grouped according to their chemical structure and representative examples from each class were selected for detailed discussion.     

Graduate students' teaching experiences improve their methodological research skills

Science, technology, engineering, and mathematics (STEM) graduate students are often encouraged to maximize their engagement with supervised research and minimize teaching obligations. However, the process of teaching students engaged in inquiry provides practice in the application of important research skills. Using a performance rubric, we compared the quality of methodological skills demonstrated in written research proposals for two groups of early career graduate students (those with both teaching and research responsibilities and those with only research responsibilities) at the beginning and end of an academic year. After statistically controlling for preexisting differences between groups, students who both taught and conducted research demonstrate significantly greater improvement in their abilities to generate testable hypotheses and design valid experiments. These results indicate that teaching experience can contribute substantially to the improvement of essential research skills.     

Excavation of a chimpanzee stone tool site in the African rainforest

Chimpanzees from the Tai forest of C?te d'Ivoire produce unintentional flaked stone assemblages at nut-cracking sites, leaving behind a record of tool use and plant consumption that is recoverable with archaeological methods. About 40 kilograms of nutshell and 4 kilograms of stone were excavated at the Panda 100 site. The data unearthed show that chimpanzees transported stones from outcrops and soils to focal points, where they used them as hammers to process foodstuff. The repeated use of activity areas led to refuse accumulation and site formation. The implications of these data for the interpretation of the earliest hominin archaeological record are explored.     

A single IGF1 allele is a major determinant of small size in dogs

The domestic dog exhibits greater diversity in body size than any other terrestrial vertebrate. We used a strategy that exploits the breed structure of dogs to investigate the genetic basis of size. First, through a genome-wide scan, we identified a major quantitative trait locus (QTL) on chromosome 15 influencing size variation within a single breed. Second, we examined genetic variation in the 15-megabase interval surrounding the QTL in small and giant breeds and found marked evidence for a selective sweep spanning a single gene (IGF1), encoding insulin-like growth factor 1. A single IGF1 single-nucleotide polymorphism haplotype is common to all small breeds and nearly absent from giant breeds, suggesting that the same causal sequence variant is a major contributor to body size in all small dogs.