Preliminary biological evaluation of some formulated feeds for P. monodon

Rice bran is widely used by fish farmers as supplementary feed while soybean cake is used both as feed and as fertilizer in fishponds. Both fish meal and shrimp head have been found acceptable as feed ingredients. However, not much is known of the acceptability and efficiency of a mixture of these ingredients as feed for Penaeus monodon larvae. Ninety 127-day old P. monodon were measured for length and weight and were randomly divided into nine aquaria each containing 20 liters of water. These were fed 'lampirong' for two months previous to the study. There were three replications for each treatment. Length, weight, and survival rates were used to compare the efficiency of the diets. Weighed amounts of pellets equivalent to 100% of the body weight were fed during the first three days and reduced to 50% thereafter. A stopwatch was used to determine the length of time that elapsed before the shrimps would approach the pellet. Ten shrimps approximately 4 months in age were placed in 10 liters of water in a 25-liter aquarium. Two grams of each pellet type were placed simultaneously on opposite sides of the aquarium. The time that elapsed from the moment the pellets sunk to the bottom up to the time that any one shrimp approached the pellets was recorded. The group fed the imported pellets gained the most. Those fed FP-2s-77 elongated faster than those fed FP-1s-77. Survival rate of those fed FP-2s-77 was 37% while those fed imported pellets was 73%. Both 1s and 2s pellets disintegrated in water easily but the imported pellets were stable even after six hours in water. The attractability test for the pellets showed that the prawns were more readily attracted to the pellets 1s and 2s than to the imported pellets.

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 .

Studies on distribution, diversity and production of macrobenthic fauna in the Chahbahar Bay

The population density, distribution, diversity and secondry production of macrobenthic fauna of the inner Chahbahar Bay were studied through bi-monthly sampling from April 1995 to March 1996. Samples were collected from water near the bottom and sediment at 14 stations inside the Bay and one reference station located outside at the entrance to the Bay. The environmental parameters Such as temperature, water depth, salinity, pH and dissolved oxygen as well as percentage silt-clay and total organic matter of the sediment were measured. The faunal population density and their distribution is discussed in relation to the environmental changes. results obtained indicated both spatial and temporal heterogeneity in faunal distribution of the Chahbahar Bay. The total of 18 groups of macrofauna were identified in all samples. Amphipods formed the dominant group (21%) followed by polychaetes (19%), gastropods (15.7%) bivalves (10.6%) and all other groups (33.7%). Seasonal changes in faunal density is shown in relation to Indian Ocean southwest monsoon,the result of which indicated lower population density during monsoon (June to September) than that of the premonsoon (February to May) and post monsoon (October to January) periods. The numerical abundance of macrobenthos varied from 10260.m2 before monsoon to 5190 m2 during monsoon season. Three dominant groups of macrofauna including polychaetes, gastropods, and bivalves were identified in all collected samples. Indices of diversity, richness and evenness were calculated for these three dominant groups. The Shannon-Weaver information index was used to describe the spatially and temporally variation in diversity of these three major faunal groups. The results exhibited lower faunal diversity during monsoon period. The annual production of two dominant macrofauna species including a species of bivalve, Nuculana acuta and a species of Cephalochordata, Branchiostoma lanceolatum were measured by using age group determination. Furthermore the mean biomass and total annual production of macrobenthic fauna were estimated for the whole studied area. The potential yield of demersal fishery resources (fish and crustacean) then estimated and worked out to be 15360 tons/year asuming 10% ecological efficiency of hypothetical pyramid from 3rd to 4th marine trophic level. Accordingly the annual exploitable demersal fishery resources for the entire Chahbahar Bay was estimated to be 7600 to 8500 tons/year by taking 50 to 55% of the total estimated potential in to account.