Aquatic ecology monitoring: water quality, fish, fish catch, sanitation and disease vectors: monitoring No. 5, 6th-9th October 2009

The survey covered by this report was undertaken between 6 th and 9th October 2009 as a follow-up on the during construction surveys of the Bujagali Hydropwer Project (BHPP). In addition to two pre-construction baseline surveys in April 2000 and April 2006, four monitoring surveys have so far been undertaken i.e. in September 2007, April 2008, April 2009 and the present one, in October 2009. The 2009 biannual monitoring surveys were conducted at an upstream and a downstream transect of the BHPP with emphasis on the following aspects: 1. water quality determinants 2. biology and ecology of fishes and food webs 3. fish stock and fish catch including economic aspects of catch and 4. sanitation/vector studies (bilharzias and river blindness)

Aquatic ecology monitoring: water quality, fish, fish catch, sanitation and disease vectors: monitoring no. 8, 6– 10th April 2011

The result reported were from a monitoring survey no.8 undertaken between 6th and 10th April 2011 during construction period of the Bujagali Hydropwer Project (BHPP). Two pre-construction, baseline surveys in April 2000 and April 2006 were conducted and so far,durin construction phase of the project, seven monitoring surveys have been undertaken i.e. in September 2007, April 2008, April 2009,October 2009, April 2010, September 2010 and the present one, in April 2011. Since 2009 biannual monitoring surveys have been conducted at an upstream and a downstream transect of the BHPP with emphasis on the following aspects: Water quality determinants Biology and ecology of fishes and food webs Fish stock and fish catch including economic aspects of catch and Sanitation/vector studies (bilharzias and river blindness)

Cloning,Expression and Sequence Analysis of A Luciferase Gene from the Chinese Firefly Pyrocoelia pygidialis

从一种来自中国日行性萤火虫(云南窗萤)发光器官mRNA中克隆、测序并表达了有功能的荧光素酶.云南窗萤荧光素酶的cDNA序列有1647个碱基,编码548个氨基酸残基.从推测得到的氨基酸序列的比对分析得出:云南窗萤的荧光素酶与来自Lampyris noctiluca,L.turkestanicus和Nyctophila cf.caucasica三种萤火虫的荧光素酶有97.8%的序列一致性.从推测得出的氨基酸序列进行系统发育分析,其结果表明:云南窗萤和Lampyris+Nyctophila聚在一起,与同属的发光强夜行性的萤火虫不形成的单系.云南窗萤荧光素酶在大肠杆菌中表达的条带大约70kDa,并且在有荧光素存在时发出黄绿色荧光.对荧光素酶的结构模拟和分析表明,云南窗萤荧光素酶基因的氨基端和羧基端结构域之间的裂沟处存在这5个多肽环,这正是从其他荧光素酶推测得到的催化荧光反应时的底物结合位点.云南窗萤和窗萤属的其他3种萤火虫的荧光素酶卡目比,有13个不同氨基酸位点,位于模拟分子结构的表面.对于这些多肽环、不刚氨基酸残基和晶体结构的进一步研究有利于解释日行和夜行性萤火虫荧光素酶的差异.

Aquatic ecology monitoring: water quality, fish, fish catch, sanitation and disease vectors: monitoring no. 10 and reservoir baseline, 23rd – 30th April 2012

The results reported on were from a monitoring survey No. 10 undertaken between 23 rd and 29th April 2012 during construction period of the Bujagali Hydropower Project (BHPP). Two pre-construction, baseline surveys in April 2000 and April 2006 were conducted and so far, during construction phase of the project, nine monitoring surveys have been undertaken i.e. in September 2007, April 2008, April 2009, October 2009, April 2010, September 2010, April 2011, September 2011and the present one, in April 2012. Since 2009 biannual monitoring surveys have been conducted at an upstream and a downstream transect of the BHPP with emphasis on the following aspects: water quality determinants biology and ecology of fishes and food webs fish stock and fish catch including economic aspects of catch and sanitation/vector studies (bilharzias and river blindness) During this survey, baseline assessment of the above mentioned studies was conducted in the reservoir behind the dam, including studies on algae, zooplankton and benthic macroinvertebrates which had been restrained since April 2008. The findings of baseline assessment of the reservoir are also contained in this report and are compared with those obtained from Transect 1(Upstream) and Transect 2 (Downstream).

Aquatic ecology monitoring, water quality, fish, fish catch, sanitation and disease vectors: monitoring no. 11, 4th – 9th September 2012

This monitoring survey No. 11 undertaken between 4th and 9th September 2012 is the second one to be conducted after completion of construction of Bujagali Hydropower Dam. Two pre-construction baseline surveys in April 2000 and April 2006 were conducted and during construction phase, eight monitoring surveys (September 2007, April 2008, April 2009, October 2009, April 2010, September 2010, April 2011, September 2011) were conducted.

Aquatic ecology monitoring: water quality, fish, fish catch, sanitation and disease vectors: monitoring No.7, 4-7th September 2010

The results reported on were from a monitoring survey No.7 undertaken between 4 th and 7th September 2010 during construction period of the Bujagali Hydropower Project (BHPP). Two pre-construction, baseline surveys in April 2000 and April 2006 were conducted and so far, during construction phase of the project, six monitoring surveys have been undertaken i.e. in September 2007, April 2008, April 2009, October 2009, April 2010 and the present one, in September 2010. Since 2009 biannual monitoring surveys have been conducted at an upstream and a downstream transect of the BHPP with emphasis on the following aspects: I. water quality determinants 2. biology and ecology of fishes and food webs 3. fish stock and fish catch including economic aspects of catch and 4. sanitation/vector studies (bilharzias and river blindness)

Aquatic ecology monitoring: water quality, fish, fish catch, sanitation and disease vectors: monitoring no.12, 25th-30th April 2013

Bujagali hydropower dam construction is now completed and a reservoir behind the dam has been created, extending all the way up to Kalange-Makwanzi, an upstream transects. During the 10th monitoring survey-April 2012, a third transect was established in the mid of the reservoir where it runs up to 30 m deep and sampled similarly as at the two original sampling transects, Kalange-Makwanzi and Buyala-Kikubamutwe for comparative purposes. This monitoring survey No. 12 undertaken between 25th and 30th April 2013 is the third one to be conducted after completion of construction of Bujagali Hydropower Dam. Two pre-construction baseline surveys in April 2000 and April 2006 were conducted and during construction phase, eight monitoring surveys (September 2007, April 2008, April 2009, October 2009, April 2010, September 2010, April 2011, September 2011) were conducted. Since 2009 biannual monitoring surveys have been conducted at an upstream and a downstream transect of the BHPP with emphasis on the following aspects: water quality determinants, biology and ecology of fishes and food webs, fish stock and fish catch including economic aspects of catch and sanitation/vector studies (bilharzias and river blindness). In the post-construction monitoring surveys, the assessments of algae, zooplankton and benthic macro-invertebrates which had been restrained since April 2008 were also included.

Evaluation of indigenous molluscicides in water against schistosomiasis vectors

Seven varieties of indigenous Phytolacca dodecwulra L'Herrit (Phytolaccaceae) were field-tried for molluscicidal potency. Varieties (U96) and (U95) collected from Kabarole and Kabale respectively were the most potent with LD90 equal to 2.54 and 6.46 mg.t-· respectively. Water bodies ranging between 4,770 and 347,510 Iitres in Kibimba rice fields were treated with up to 50mg.t-· Snails kills were monitored every three months and 92 - 100% mortality rates were realized. HPLC fingerprints revealed the two P. dodecandra varieties to contain highest concentration of the active principle, oleanoglycotoxin- A or lemmatoxin - A.

Ultrasonic imaging of 3D displacement vectors using a simulated 2D array and beamsteering.

Most quasi-static ultrasound elastography methods image only the axial strain, derived from displacements measured in the direction of ultrasound propagation. In other directions, the beam lacks high resolution phase information and displacement estimation is therefore less precise. However, these estimates can be improved by steering the ultrasound beam through multiple angles and combining displacements measured along the different beam directions. Previously, beamsteering has only considered the 2D case to improve the lateral displacement estimates. In this paper, we extend this to 3D using a simulated 2D array to steer both laterally and elevationally in order to estimate the full 3D displacement vector over a volume. The method is tested on simulated and phantom data using a simulated 6-10MHz array, and the precision of displacement estimation is measured with and without beamsteering. In simulations, we found a statistically significant improvement in the precision of lateral and elevational displacement estimates: lateral precision 35.69μm unsteered, 3.70μm steered; elevational precision 38.67μm unsteered, 3.64μm steered. Similar results were found in the phantom data: lateral precision 26.51μm unsteered, 5.78μm steered; elevational precision 28.92μm unsteered, 11.87μm steered. We conclude that volumetric 3D beamsteering improves the precision of lateral and elevational displacement estimates.

Ultrasonic imaging of 3D displacement vectors using a simulated 2D array and beamsteering

Most quasi-static ultrasound elastography methods image only the axial strain, derived from displacements measured in the direction of ultrasound propagation. In other directions, the beam lacks high resolution phase information and displacement estimation is therefore less precise. However, these estimates can be improved by steering the ultrasound beam through multiple angles and combining displacements measured along the different beam directions. Previously, beamsteering has only considered the 2D case to improve the lateral displacement estimates. In this paper, we extend this to 3D using a simulated 2D array to steer both laterally and elevationally in order to estimate the full 3D displacement vector over a volume. The method is tested on simulated and phantom data using a simulated 6-10 MHz array, and the precision of displacement estimation is measured with and without beamsteering. In simulations, we found a statistically significant improvement in the precision of lateral and elevational displacement estimates: lateral precision 35.69 μm unsteered, 3.70 μm steered; elevational precision 38.67 μm unsteered, 3.64 μm steered. Similar results were found in the phantom data: lateral precision 26.51 μm unsteered, 5.78 μm steered; elevational precision 28.92 μm unsteered, 11.87 μm steered. We conclude that volumetric 3D beamsteering improves the precision of lateral and elevational displacement estimates. © 2012 Elsevier B.V. All rights reserved.

Hybrid cytomegalovirus-U6 promoter-based plasmid vectors improve efficiency of RNA interference in zebrafish

Short hairpin RNA (shRNA) directed by RNA polymerase III (Pol III) or Pol II promoter was shown to be capable of silencing gene expression, which should permit analyses of gene functions or as a potential therapeutic tool. However, the inhibitory effect of shRNA remains problematic in fish. We demonstrated that silencing efficiency by shRNA produced from the hybrid construct composed of the CMV enhancer or entire CMV promoter placed immediately upstream of a U6 promoter. When tested the exogenous gene, silencing of an enhanced green fluorescent protein (EGFP) target gene was 89.18 +/- 5.06% for CMVE-U6 promoter group and 88.26 +/- 6.46% for CMV-U6 promoter group. To test the hybrid promoters driving shRNA efficiency against an endogenous gene, we used shRNA against no tail (NTL) gene. When vectorized in the zebrafish, the hybrid constructs strongly repressed NTL gene expression. The NTL phenotype occupied 52.09 +/- 3.06% and 51.56 +/- 3.68% for CMVE-U6 promoter and CMV-U6 promoter groups, respectively. The NTL gene expression reduced 82.17 +/- 2.96% for CMVE-U6 promoter group and 83.06 +/- 2.38% for CMV-U6 promoter group. We concluded that the CMV enhancer or entire CMV promoter locating upstream of the U6-promoter could significantly improve inhibitory effect induced by the shRNA for both exogenous and endogenous genes compared with the CMV promoter or U6 promoter alone. In contrast, the two hybrid promoter constructs had similar effects on driving shRNA.

Integration of double-fluorescence expression vectors into zebrafish genome for the selection of site-directed knockout/knockin

Production of zebrafish by modifying endogenous growth hormone (GH) gene through homologous recombination is described here. We first constructed the targeting vectors pGHT1.7k and pGHT2.8k, which were used for the knockout/knockin of the endogenous GH gene of zebrafish, and injected these two vectors into the embryos of zebrafish. Overall, the rate of targeted integration with the characteristic of germ line transmission in zebrafish was 1.7x10(-6). In one experimental patch, the integrating efficiency of pGHT2.8k was higher than that of pGHT1.7k, but the lethal effect of pGHT2.8k was stronger than that of pGHT1.7k. The clones with the correct integration of target genes were identified by a simple screening procedure based on green fluorescent protein (GFP) and RFP dual selection, which corresponded to homologous recombination and random insertion, respectively. The potential homologous recombination zebrafish was further bred to produce a heterozygous F-1 generation, selected based on the presence of GFP. The potential targeted integration of exogenous GH genes into a zebrafish genome at the P-0 generation was further verified by polymerase chain reaction and Southern blot analysis. Approximately 2.5% of potential founder knockout and knockin zebrafish had the characteristic of germ line transmission. In this study, we developed an efficient method for producing the targeted gene modification in zebrafish for future studies on genetic modifications and gene functions using this model organism.

The effect of nocodazole on the transfection efficiency of lipid-bilayer coated gold nanoparticles

Nonviral vectors are safer than viral systems for gene therapy applications. However, the limited efficacy always prevents their being widely used in clinical practice. Aside from searching new gene nonviral vectors, many researchers focus on finding out new substances to improve the transfection efficiency of existent vectors. In this work, we found a transfection enhancer, nocodazole (NCZ), for dimethyldioctadecylammonium (DODAB, a cationic lipid) bilayer coated gold nanoparticles (AuNPs) mediated gene delivery. It was found that NCZ produces 3-fold transfection enhancement to HEK 293T cells assessed by flow cytometry (FCM). The result was further confirmed by luciferase assay, in which NCZ induced more than 5 times improvement in transfection efficiency after 48 h of transfection. The results from the inductively coupled plasma mass spectrometry (ICP-MS) and FCM showed that NCZ did not affect the internalization of DODAB-AuNPs/DNA complexes. The trafficking of the complexes by transmission electron microscopy (TEM) indicated that the interrupted transportation of the complexes to the lysosomes contributed greatly to the transfection enhancement.

Multi-armed poly(L-glutamic acid)-graft-oligoethylenimine copolymers as efficient nonviral gene delivery vectors

Background The application of polyethylenimine (PEI) in gene delivery has been severely limited by significant cytotoxicity that results from a nondegradable methylene backbone and high cationic charge density. It is therefore necessary to develop novel biodegradable PEI derivates for low-toxic, highly efficient gene delivery.Methods A series of novel cationic copolymers with various charge density were designed and synthesized by grafting different kinds of oligoethylenimine (OEI) onto a determinate multi-armed poly(L-glutamic acid) backbone. The molecular structures of multi-armed poly(L-glutamic acid)-graft-OEI (MP-g-OEI) copolymers were characterized using nuclear magnetic resonance, viscosimetry and gel permeation chromatography. Moreover, the MP-g-OEI/DNA complexes were measured by a gel retardation assay, dynamic light scattering and atomic force microscopy to determine DNA binding ability, particle size, zeta potential, complex formation and shape, respectively. MP-g-OEI copolymers were also evaluated in Chinese hamster ovary and human embryonic kidney-293 cells for their cytotoxicity and transfection efficiency.