Growth response and survival of F1 hybrid fry of Heterobranchus longifilis and Clarias gariepinus reared in glass aquaria and plastic basins

The growth response of F1 hybrid fry of female Heterobranchus longifilis and male Clarias gariepinus were investigated under laboratory conditions in glass aquaria glass tanks and plastic basins. The larvae were produced artificially after inducement with Ovarptim. The hatching percentage was very high. Weekly mean length and weight were monitored for 6 weeks. The average length increase was higher in aquaria glass tanks than in plastic basins. However, there were depressed and irregular weight increases in both types of rearing troughs while significant weight increase (P<0.05) was recorded at week 6 in the plastic basin. Generally, the growth rate and survival in both containers were not significantly different

Monoculture of three native air breathing fishes in homestead ditches and plastic barrels

Six on-farm trials were conducted from 1 August to 23 November 2004 in two different environments such as homestead ditches (10 to 17 square meters) and plastic barrels (240 liters) to develop techniques for nonoculture [sic] of climbing perch, Anabas testudineus, stinging catfish, Heteropneustes fossilis and walking catfish, Clarias batrachus for poor and landless people who have no access to pond. Stocking density for ditch was 10 fry/square meter while that for barrel was 20 fry/cubic meter. The fishes were fed with 3-test diets viz. low-cost formulated feed (rice bran 20%, wheat meal 10%, mustered [sic] oil cake 35%, poultry offal 35%), live foods (chopped snails and clams), and a commercial feed (Saudi-Bangla feed, starter 3: first month and grower-1: subsequent two months) and designated as T1, T2 and T3, respectively. Feeding rate was the same in all the treatments viz. 10% of body weight (first two months), 8% (third month) and 6% (fourth month). T1 and T2 had three replications while T3 had two replications. Water temperature was recorded weekly while fish growth was monitored monthly. After 4 months' rearing, H. fossilis and C. batrachus in ditches and barrels attained higher average weight in T2 followed by T3 and T1 while A. testudineus in barrels also attained higher average weight in T2. The variation in net weight gain by A. testudineus in ditch fed test diets T2 and T3 was not significantly different (P>0.05) though the net gain in both T2 and T3 was significantly (P<0.05) higher than that of T1. The yield of climbing perch as obtained from T1, T2 and T3 was 988, 1136 and 1185 kg/ha, respectively while that stinging catfish was 395, 242 and 444 kg/ha and walking catfish was 1605, 2,099 and 1,654 kg/ha respectively. All the three species showed significantly lower growth rate in barrels than in ditches.

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 .