Polyphosphate kinase genes from full-scale activated sludge plants

The performance of enhanced biological phosphorus removal (EBPR) wastewater treatment processes depends on the presence of bacteria that accumulate large quantities of polyphosphate. One such group of bacteria has been identified and named Candidatus Accumulibacter phosphatis. Accumulibacter-like bacteria are abundant in many EBPR plants, but not much is known about their community or population ecology. In this study, we used the polyphosphate kinase gene (ppk1) as a high-resolution genetic marker to study population structure in activated sludge. Ppk1 genes were amplified from samples collected from full-scale wastewater treatment plants of different configurations. Clone libraries were constructed using primers targeting highly conserved regions of ppk1, to retrieve these genes from activated sludge plants that did, and did not, perform EBPR. Comparative sequence analysis revealed that ppk1 fragments were retrieved from organisms affiliated with the Accumulibacter cluster from EBPR plants but not from a plant that did not perform EBPR. A new set of more specific primers was designed and validated to amplify a 1,100 bp ppk1 fragment from Accumulibacter-like bacteria. Our results suggest that the Accumulibacter cluster has finer-scale architecture than previously revealed by 16S ribosomal RNA-based analyses. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

A bioreactor model system specifically designed for <Emphasis Type="Italic">Tetrahymena</Emphasis> growth and cholesterol removal from milk

This work describes the configuration and operation of a bioreactor system especially designed for Tetrahymena cultivation and its use for milk improvement, particularly cholesterol elimination by the action of this cell. An advantage of the proposed method is the re-use of the growth medium; thus, the medium is used twice to provide two batches of Tetrahymena biomass without the need of further inoculation. This makes the procedure of producing the cell biomass faster and more economical. Cells are concentrated in the culture vessels by sedimentation at room temperature and then transferred to milk suspensions, where they can further grow for at least one generation with the benefit of reducing steeply cholesterol level. Milk treated according to this process is separated from the biomass by centrifugation. Under these conditions, less than 5% of the cells remain in the milk, and cholesterol elimination amounts to 75 +/- 10% of that initially present. No changes in sensorial properties of the milk, such as clotting or butyric odor, were observed as a result of this treatment. In addition, the bioreactor allows the aseptic recovery of the spent growth medium, which contains diverse enzymes of interest, and the cell pellets, to exploit particular lipids like phosphonolipids, abundant poly-unsaturated fatty acids and co-enzyme Q(8).     

Estimating Wind Turbine-Caused Bird Mortality

ABSTRACT Mortality estimates are needed of birds and bats killed by wind turbines because wind power generation is rapidly expanding worldwide. A mortality estimate is based on the number of fatalities assumed caused by wind turbines and found during periodic searches, plus the estimated number not found. The 2 most commonly used estimators adjust mortality estimates by rates of searcher detection and scavenger removal of carcasses. However, searcher detection trials can be biased by the species used in the trial, the number volitionally placed for a given fatality search, and the disposition of the carcass on the ground. Scavenger removal trials can be biased by the metric representing removal rate, the number of carcasses placed at once, the duration of the trial, species used, whether carcasses were frozen, whether carcasses included injuries consistent with wind turbine collisions, season, distance from the wind turbines, and general location. I summarized searcher detection rates among reported trials, and I developed models to predict the proportion of carcasses remaining since the last fatality search. The summaries I present can be used to adjust previous and future estimates of mortality to improve comparability. I also identify research directions to better understand these and other adjustments needed to compare mortality estimates among wind farms.     

Potential application of cyclic lipopeptide biosurfactants produced by Bacillus subtilis strains in laundry detergent formulations

AIMS: Crude cyclic lipopeptide (CLP) biosurfactants from two Bacillus subtilis strains (DM-03 and DM-04) were studied for their compatibility and stability with some locally available commercial laundry detergents. METHODS AND RESULTS: CLP biosurfactants from both B. subtilis strains were stable over the pH range of 7.0-12.0, and heating them at 80 degrees C for 60 min did not result in any loss of their surface-active property. Crude CLP biosurfactants showed good emulsion formation capability with vegetable oils, and demonstrated excellent compatibility and stability with all the tested laundry detergents. CONCLUSION: CLP biosurfactants from B. subtilis strains act additively with other components of the detergents to further improve the wash quality of detergents. The thermal resistance and extreme alkaline pH stability of B. subtilis CLP biosurfactants favour their inclusion in laundry detergent formulations. SIGNIFICANCE AND IMPACT OF THE STUDY: This study has great significance because it is already known that microbial biosurfactants are considered safer alternative to chemical or synthetic surfactants owing to lower toxicity, ease of biodegradability and low ecological impact. The present study provides further evidence that CLP biosurfactants from B. subtilis strains can be employed in laundry detergents.     

Army ants in four forests: geographic variation in raid rates and species composition

1. The New World army ants are top predators in the litter of tropical forest, but no comprehensive studies exist on variation in assemblage-wide activity and species composition. We used standardized protocols to estimate foraging raid rates and species composition of army ant communities in four Neotropical forests. The study sites spanned approximately 10 degrees latitude, with two sites each in Central and South America. 2. We recorded a total of 22 species of army ants. The four sites varied in observed and estimated species richness. Species overlap was highest between the Central American sites, and lowest between the South American sites. 3. Raid activity varied significantly among sites. Raid activity per kilometre of trail walks was over four times higher at the most active site (Sta. Maria, Venezuela) than at the least active site (Barro Colorado Island, Panama). Furthermore, each site showed a different diel pattern of activity. For example, raid activity was higher during daylight hours in Costa Rica, and higher at night in Venezuela. Raid activity relationships with ambient temperature also varied significantly among sites. 4. The overall rate of army ant raids passing through 1 m(2) plots was 0.73 raids per day, but varied among sites, from 0 raids per day (Panama) to 1.2 raids per day (Venezuela). 5. Primarily subterranean species were significantly more abundant in Venezuela, and above-ground foragers that form large swarm fronts were least abundant in Panama. The site heterogeneity in species abundance and diel activity patterns has implications for army ant symbionts, including ant-following birds, and for the animals hunted by these top predators.     

珠海市陆生天然次生林优势种的生态位

在珠海市10个陆生次生林群落中设置了112个10m×10m样方,用于珠海市次生林群落23个优势种的生态位宽度和生态位重叠的研究。结果表明:珠海次生林群落23个优势种的Shannon-Wiener生态位宽度和Levins生态位变度值范围为0.4049～0.7736和0.0551～0.3086,生态位宽度较大的种类分别是山油柑(Acronychia pedunculata)、大头茶(Gordonia axillaris)、银柴(Aporosa dioica)、小盘木(Microdesmis casearifolia)、白颜树(Giron-niera subaequalis)和假苹婆(Sterculia lanceolata);生态位相似性比例值为0～0.3810,银柴与阴香(Cinnamomum burmanii)的生态位相似性比例值最高;山油柑与另外22个优势种均存在不同程度的生态位重叠;大头茶只与少数优势种存在生态位重叠;另有20.6%的种对之间不存在生态位重叠。建议使用山油柑、大头茶、白颜树和黄桐(Endospermum chinense)等生态位宽度较高、分布广泛、适应力强的树种,进行森林恢复与人工林改造。     

脱氮菌株P6的分离鉴定及其处理氨氮废水的试验研究

目的：从污水处理池的污泥中分离得到去除氨氮效果较好的菌株,对其进行分类鉴定和去除氨氮的最佳条件的研究。方法：根据维诺格拉斯基柱法分离出几种纯菌株,选取其中脱氮效果较好的一株,经形态观察和生理生化实验,鉴定其分类地位。研究不同培养条件对该菌生长的影响,用滴定法测定不同条件下其对氨氮的去除率。结果：获得一脱氮菌株P6,初步推测该菌为产碱杆菌。其最适生长条件为：温度29℃,初始pH值7.5以及摇床（100r/min）培养,且在温度29℃、废水的初始pH值11、接种量与废水体积的比例为1.2g/150mL、添加乙酸钠浓度为0.1%、废水的氨氮浓度为600mg/L和摇床（100r/min）的条件下,3d后该菌对氨氮的去除率达到45%。结论：P6菌有一定的去除氨氮能力,具有应用价值。     

Dominance Relationship between Two Self-Incompatible<Emphasis Type="Italic">Brassica campestris</Emphasis> Haplotypes in Response to CO<Subscript>2</Subscript> Gas

InBrassica, self-incompatibility (SI) can be overcome by CO₂ application, an effective method for obtaining numerous inbred lines for F, commercial seed. We previously reported two different S-alleles ofBrassica campestris, S⁷³³ and S⁷³⁴, with extremely different degrees of susceptibility to this gas. In the current study, we raised a cross-population between those two genetic lines, and analyzed their reaction level of self-incompatibility to CO₂ (RLSICO₂). Here, all 40 of our progeny from the F₁ cross-population were susceptible, maintaining high values of RLSICO₂. This suggests that the susceptible line, S⁷³⁴, is dominant to the insusceptible line, S⁷³³. We also generated an F₂ selfing-population of each crossed progeny, S⁷³³♀ S⁷³⁴♂ and S⁷³³♂ S⁷³⁴♀, to assess the RLSICO₂ of each individual. PCR-RFLP analysis was performed to determine the S-genotype of the F₂ population. The S⁷³⁴ allele segregated in a theoretical ratio of the dominant trait, and the RLSICO₂ was consistent with the dominance relationship. Therefore, we have now demonstrated that high RLSICO₂ in β.campestris is controlled by a dominant gene. Both authors contributed equally to this work     

Partitioning and inactivation of viruses by the caprylic acid precipitation followed by a terminal pasteurization in the manufacturing process of horse immunoglobulins

Caprylic acid (octanoic acid), has been used for over 50 years as a stabilizer of human albumin during pasteurization. In addition caprylic acid is of great interest, by providing the advantage of purifying mammalian immunoglobulins and clearing viruses infectivity in a single step. Exploiting these two properties, we sequentially used the caprylic acid precipitation and the pasteurization to purify horse hyperimmune globulins used in the manufacturing of Sérocytol. To evaluate the effectiveness of the process for the removal/inactivation of viruses, spiking studies were carried out for each dedicated step. Bovine viral diarrhoea virus (BVDV), pseudorabies virus (PRV), encephalomyocarditis virus (EMCV) and minute virus of mice (MVM) were used for the virological validation. Our data show that the treatment with caprylic acid 5% (v/v) can effectively be used as well to purify or to ensure viral safety of immunoglobulins. Caprylic acid precipitation was very efficient in removing and/or inactivating enveloped viruses (PRV, BVDV) and moderately efficient against non-enveloped viruses (MVM, ECMV). However the combination with the pasteurization ensured an efficient protection against both enveloped and non-enveloped viruses. So that viruses surviving to the caprylic acid precipitation will be neutralized by pasteurization. Significant log reduction were achieved > or =9 log(10) for enveloped viruses and 4 log(10) for non-enveloped viruses, providing the evidence of a margin of viral safety achieved by our manufacturing process. Its a simple and non-expensive manufacturing process of immunoglobulins easily validated that we have adapted to a large production scale with a programmable operating system.     

Sludge wash-out as strategy for Anammox process start-up

In this study sludge wash-out was evaluated as a strategy to start-up the Anammox process in order to establish it in a shorter period of time. Sludge from a domestic wastewater treatment plant (WTP) was used to seed two (RI and RII) anaerobic sequencing batch reactors (ASBR). During the start-up period RI was operated as a continuous stirred tank reactor (CSTR) using a dilution rate of 0.2 d⁻¹, which promoted the sludge wash-out. After this period, the remaining sludge was retained in the reactor. The reactor RII was operated as an ASBR throughout the study period with a high cell retention. The performance of the two reactors in terms of nitrogen removal was compared over a period of 380 days. During the last RI operation phase the specific nitrogen removal rate increased exponentially, attaining values of 85 mg N/g TSS d. However, a rate of 190 mg N/g TSS d in the batch test under optimal conditions was achieved. The specific nitrogen removal rate remained almost constant for RII with a mean value of 6 mg N/g TSS d being observed during the operation period. The rate for the RII batch test was 20 mg N/g TSS d. These results confirm that the higher total suspended solids (TSS) in RII (reactor with high cell retention) was not effective in terms of N removal improvement. Anammox-like bacteria were found using fluorescence in situ hybridization (FISH) in reactor RI after 225 days and a new Anammox species was identified.