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 .

Study ectoparasite infestation of 10 common imported freshwater ornamental fish species

In the present study, ectoparasite infestation of common freshwater ornamental fish species imported into Iran including: Dwarf gourami (Colisa lalia), Angelfish (Pterophyllum scalare), Oscar (Astronatus ocellatus), Red eye Tetra ( Moenkhansia Sanctaefilomenae), Barb( Capoeta tetrazona), Arowana (Osteoglossum bicirrhosum), Goldfish(Carassius auratus), Red fin Shark (Lebeo elythrurus, Catfish (Hypostomus plecostomus) and Pangasius sutchi ( Pangasius hypophthalmus) from May until April was investigated.A total of 6 specimens sample of each fish species) randomly were obtained and taken alive to the lab. After observing gill and skin wet smear under the microscope , gill was dissected and examined carefully .Photo and films were taken from the isolated parasites and parasite identification was performed according to Yamaguti AP1), Bychowsky (■Y).From a total of examined fishes 1\ ► sample (YY.V.) were parasitized .Parasite groups that were observed consisting ciliated protozoan myxosporidian , monogenean Digenean metacercaria and crustacean .The only non-parasitized fish was Barb. Monogenea Trianchoratus sp. , Cleidodiscus sp. , Ancylodiscoides sp. , Thaparocleidus sp. , Centrocestus formosanus metacercaria and Myxobolus longisporus report for the first time in ornamental fish which imported from Southeast of Asia to Iran. According to the results, it seems that severe quarantine and sanitary rules are necessary.

Some major unsolved aspects of the dynamics of African fisheries as related to questions of rational development and management

Studies of fish and fisheries in Africa fall in to four phases: the period of fisheries expeditions, ecological investigations, the development phase, and the period of mechanized exploitation. There is need to establish the taxonomic status and ecology of the varied components of the potentially important Haplochromis in Lake Victoria. A comprehensive study of their bionomics and life history, population structure, natality, recruitment and mortality coefficients should be undertaken. Emphasis lo be laid on the study of the ecology, especially breeding behaviour of the economically important c1upeids (Stolothrissa tanganciae and Limnothrissa miodon), in Lake Tanganyika. A comprehensive investigation into the migratory and shoaling behaviour of the Lake Victoria Tilapia to be initiated. Pre-impoundment studies to be undertaken to assess effects of hydroelectric projects of fisheries. Studies on parasites of economically important fishes to be stepped up to assess pathological effects and the biological basis for their control. The role of predators, e.g., Hydrocyon, Lates and Micropterus salmoides in commercial fish populations should be evaluated, and the knowledge gaincd used to effectively manage the fisheries in favour of the more desirable fish stocks.

Biological observations in Indian mackerel, Rastrelliger kanagurta (Cuvier) 1816, (Pisces: Scombridae) from East African waters

The present study was under taken to provide further and more detailed information on the apparent seasonal and relative abundance of the species, food and feeding habits. Spawning season and size composition. The incidence of parasites, in relation to the month of the year and the fish length, was also examined.

Helminth fauna of Oreochromis leucostictus (Pisces: Cichlidae) from a tropical lake, Lake Naivasha, Kenya

The helminth fauna of Oreochromis leucosticus (Trewavas) from Lake Naivasha is described. The study was carried out on 1,050 gillnetted fish caught over a period of 24 months at different sites within the lake. Oreochromis leucosticus which is a commercially important fish was observed to be a host of four helminth parasites. These were an acanthocephalan polyacanthorhynchus kenyensis (Schmidt and Canaris), a nematode contracaecum sp.. a trematode clinostomum sp. and a dilepidid cestode Amirthalingamia sp. Seasonal variation in prevalence, intensity of infection, organs infected and the variation in infection with the size, sex and body condition of the host are described. There was no seasonal variation in prevalence but intensity of infection was observed to increase with the age of the fish. Although male fish were observed to be more heavily infected, the parasites did not seem to have any significant effect on the body condition of the fish.