Anadromous fish as marine nutrient vectors

The tidal freshwater of Virginia supports anadromous herring (Alosa spp.) spawning runs in the spring; however, their importance as nutrient delivery vectors to the freshwater fish food web remains unknown. The stable isotope signatures of fishes from 21 species and four different guilds (predators, carnivores, generalists, and planktivores) were examined in this study to test the hypothesis that marine derived nutrients (MDNs) brought by anadromous fish would be traced into the guilds that incorporated them. Spawning anadromous fish were 13C and 34S-enriched (δ13C and δ34S of approximately 18‰ and 17.7‰, respectively) relative to resident freshwater fish. Of the guilds examined, only predators showed 13C and 34S-enrichment similar to the anadromous fish; however, some generalist catfish also showed enriched signatures. Specific fatty acid δ13C signatures for gizzard shad (Dorosoma cepedianum), blue catfish (Ictalurus furcatus), and alewife (Alosa pseudoharengus), show a 10‰ range among fishes, clearly reflecting isotopically distinct dietary sources. The δ13C and δ34S distribution and range among the freshwater fishes suggest that both autochthonous and allochthonous (terrestrial C3 photosynthetic production and MDN) nutrient sources are important to the tidal freshwater fish community.

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 and expression of zebra fish ferroportin and investigating its influence on anemia

Iron is required for many microbes and pathogens for their survival and proliferation including Leishmania which cause leishmaniasis. Leishmaniasis is an increasingly serious infectious disease with a wide spectrum of clinical manifestations. These range from localized cutaneous leishmaniasis (CL) lesions to a lethal visceral form. Certain strains such as BALB/c mice fail to control L. major infection and develop progressive lesions and systemic disease. These mice are thought to be a model of non-healing forms of the human disease such as kala-azar or diffuse cutaneous leishmaniasis. Progression of disease in BALB/c mice has been associated with the anemia, in last days of their survival, the progressive anemia is considered to be one of the reasons of their death. Ferroportin (Fpn), a key regulator of iron homeostasis is a conserved membrane protein that exports iron across the duodenal enterocytes as well as macrophages and hepatocytes into the blood circulation. Fpn has also critical influence on survival and proliferation of many microorganisms whose growth is dependent upon iron, thus preparation of Fpn is needed to study the role of iron in immune responses and pathogenesis of micoorganisms. To prepare and characterize a recombinant ferroportin, total RNA was extracted from Indian zebrafish duodenum, and used to synthesize cDNA by RT-PCR. PCR product was first cloned in Topo TA vector and then subcloned into the GFP expression vector pEGFP–N1. The final resulted plasmid (pEGFP-ZFpn) was used for expression of FPN-EGFP protein in Hek 293T cells. The expression was confirmed by fluorescence microscopy and flow cytometery. Recombinant Fpn was further characterized by submission of its predicted amino acid sequences to the TMHMM V2.0 prediction server (hidden Markov model), NetOGlyc 3.1 server and NetNGlyc 3.1 server. Data emphasised that obtained Fpn from indian zebrafish contained eight transmembrane domains with N- and C-termini inside the cytoplasm and harboured 78 mucin-type glycosylated amino acid. The results indicate that the prepared and characterized recombinant Fpn protein has no membrane topology difference compared to other Fpn described by other researcher. Our next aim was to deliver recombinant plasmid (pEGFP-ZFpn) to entrocyte cells. However, naked therapeutic genes are rapidly degraded by nucleases, showing poor cellular uptake, nonspecificity to the target cells, and low transfection efficiency. The development of safe and efficient gene carriers is one of the prerequisites for the success of gene therapy. Chitosan and alginate 139 polymers were used for oral gene carrier because of their biodegradability, biocompatibility and their mucoadhesive and permeability-enhancing properties in the gut. Nanoparticles comprising Alginate/Chitosan polymers were prepared by pregel preparation method. The resulting nanoparticles had a loading efficiency of 95% and average size of 188 nm as confirmed by PCS method and SEM images had showed spherical particles. BALB/c mice were divided to three groups. The first and second group were fed with chitosan/alginate nanoparticles containing the pEGFP-ZFpn and pEGFP plasmid, respectively (30 μgr/mice) and the third group (control) didn’t get any nanoparticles. The result showed BALB/c mice infected by L.major, resulted in higher hematocryte and iron level in pEGFP-ZFpn fed mice than that in other groups. Consentration of cytokines determined by ELISA showed lower levels of IL-4 and IL-10 and higher levels of IFN-γ/IL-4 and IFN-γ/IL-10 ratios in pEGFP-ZFpn fed mice than that in other groups. Morover more limited increase of footpad thickness and significant reduction of viable parasites in lymph node was seen in pEGFP-ZFpn fed mice. The results showed the first group exhibited a highr hematocryte and iron compared to the other groups. These data strongly suggests the in vivo administration of chitosan/alginate nanoparticles containing pEGFP-ZFpn suppress Th2 response and may be used to control the leishmaniasis .

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.

Aquatic ecology monitoring: water quality, fish, fish catch, sanitation and disease vectors: monitoring No.9, 9th – 12th September 2011

The results reported on were from a monitoring survey No. 9 undertaken between 9th and 12th September 2011 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, eight monitoring surveys have been undertaken i.e. in September 2007, April 2008, April 2009, October 2009, April 2010, September 2010, April 2011 and the present one, in September 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)in addition to the above mentioned studies, a soil pH survey was undertaken on 15th October 2011 in the area behind the reservoir whose filling started a week earlier. The findings of pH status in the catchment of the dam are also contained in this report.