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 .

Tidal oscillations at the head of Monterey Submarine Canyon and their relation to oceanographic sampling and the circulation of water in Monterey Bay. Annual report, Part 6, September 1972

During a 25-hour hydrographic times series at two stations near the head of Monterey Submarine Canyon, an internal tide was observed with an amplitude of 80 to 115 m in water depths of 120 and 220 m respectively. These large oscillations produced daily variations in hydrographic and chemical parameters that were of the same magnitude as seasonal variations in Monterey Bay. Computed velocities associated with the internal tide were on the order of 10 em/sec, and this tidally induced circulation may have a significant role in the exchange of deep water between Monterey Submarine Canyon and the open ocean. (PDF contains 49 pages)

Modelling of the Subarctic North Pacific Circulation (Report of Working Group 7)

This review focuses on modelling ocean circulation and its variability in the subarctic North Pacific; it addresses issues specific to that region, and not the subject of ocean modelling in general. The performance of existing models is assessed in relation to observations in the upper ocean, intermediate waters and deep/abyssal waters. (PDF contains 87 pages)

The ontogeny of haematopoiesis in the marine teleost Leiostomus xanthurus and a comparison of the site of initial haematopoiesis with Opsanus tau

The ontogeny of haematopoiesis in the perciform fish, spot Leiostomus xanthurus, differed from that reported as the norm for fishes, as exemplified by the cypriniform zebrafish Danio rerio, and observed in the batrachoidiform oyster toadfish Opsanus tau. Erythropoiesis in spot was first evident in the head kidney of yolk-sac larvae 3 days after hatching (DAH). No embryonic intermediate cell mass (ICM) of primitive stem cells or blood islands on the yolk were apparent within embryos. Erythrocytes were first evident in circulation near the completion of yolk absorption, c. 5 DAH, when larvae were c. 20 mm notochord length (LN). Erythrocyte abundance increased rapidly with larval development for c. 14 to 16 DAH, then became highly variable following changes in cardiac chamber morphology and volume. Erythrocytic haemoglobin (Hb) was not detected within whole larvae until they were 12 DAH or c. 31 mm LN, well after yolk and oil-globule absorption. The Hb was not quantified until larvae were >47 DAH or >7 mm standard length. The delayed appearance of erythrocytes and Hb in spot was similar to that reported for other marine fishes with small embryos and larvae. In oyster toadfish, a marine teleost that exhibits large embryos and larvae, the ICM and Hb were first evident in two bilateral slips of erythropoietic tissue in the embryos, c. 5 days after fertilization. Soon thereafter, erythrocytes were evident in the heart, and peripheral and vitelline circulation. Initial haematopoiesis in oyster toadfish conformed with that described for zebrafish. While the genes that code for the development of haematopoiesis are conserved among vertebrates, gene expression lacks phylogenetic pattern among fishes and appears to conform more closely with phenotypic expression related to physiological and ecological influences of overall body size and environmental oxygen availability.

Blood lactate in yellowfin tuna, Neothunnus macropterus, and skipjack, Katsuwonus pelamis, following capture and tagging

ENGLISH: The staff of the Inter-American Tropical Tuna Commission for several years has been investigating the life history, population structure, behavior and ecology of the yellowfin tuna, Neothunnus macropterus, and the skipjack, Katsuwonus pelamis, in the Eastern Tropical Pacific Ocean. The tagging and subsequent recovery of these tropical tunas, to provide information on population structure, migrations, mortality rates and growth rates, are important aspects of these investigations. Broadhead (1959) and Schaefer, Chatwin and Broadhead (1961) emphasize the many difficulties involved in tagging these extremely active yet delicate fish and give considerable evidence to suggest that tagging mortality is high, perhaps as great as 60 to 80 per cent. The latter authors suggest that the rather high mortality at tagging is related to the effects of hyperactivity brought about by the tagging operation. SPANISH: El personal de la Comisión Interamericana del Atún Tropical ha estado investigando durante varios años la historia natural, la estructura de la población, los hábitos y la ecología del atún aleta amarilla, Neothunnus macropterus, y del barrilete, Katsuwonus pelamis, en el Océano Pacífico Oriental Tropical. La marcación y el subsiguiente recobro de estos atunes tropicales, lo que da información sobre la estructura de la población, los movimientos migratorios y las tasas de crecimiento y de mortalidad, son importantes aspectos de estas investigaciones. Broadhead (1959) y Schaefer, Chatwin y Broadhead (1961) destacan las muchas dificultades que hay para marcar estos peces activos en extremo pero delicados, y proporcionan considerable evidencia que sugiere que la mortalidad por la marcación es bastante alta, siendo quizás de 60 a 80 por ciento. Los autores citados sugieren que esta elevada mortalidad por la marcación está relacionada con los efectos de la hiperactividad producida por la operación de marcación.

Muscle glycogen and blood lactate in yellowfin tuna, Thunnus albacares, and skipjack, Katsuwonus pelamis, following capture and tagging

ENGLISH: Tagging and the recovery of tagged yellowfin (Thunnus albacares) and skipjack (Katsuwonus pelamis) tunas are important aspects of the investigations conducted by the Inter-American Tropical Tuna Commission in the Eastern Tropical Pacific Ocean. The results of the tagging program provide information on population structures, migrations, mortality rates and growth rates of these two species. The present experimental program was undertaken to study the relationship between muscular fatigue and high tagging mortalities in yellowfin and skipjack. SPANISH: La marcación del atún aleta amarilla (Thunnus albacares) y del barrilete (Katsuwonus pelamis), y el recobro de estos atunes marcados, son aspectos importantes de la investigación que efectúa la Comisión Interamericana del Atún Tropical en el Océano Pacífico Oriental Tropical. Los resultados del programa de marcación proporcionan información sobre la estructura de las poblaciones, migraciones, tasas de mortalidad y tasas de crecimiento de estas dos especies. El programa experimental presente fue emprendido para estudiar la relación entre la fatiga muscular y la alta mortalidad causada por la marcación en el atún aleta amarilla y el barrilete. (PDF contains 52 pages.)

Effects of fish-meal, cow blood-meal, and sorghum diets on food utilization and growth of cage cultured Sarotherodon niloticus

The growth responses and feed utilization of Sarotherodon niloticus held in metal cages in a pond and fed diets containing fish-meal, cow blood-meal or sorghum was studied. Results indicate that the best growth, feed conversion and protein efficiency ratio were obtained with the diet containing 60% fish-meal. The growth performance of fish on 40% fish-meal, and 40% and 60% blood meal were not significantly different, and were quite close to the performance with 60% fish-meal. The growth and food utilization of fish on 84% sorghum diet was significantly lower than the rest. The Caged fish without supplemental feeding had a light gain in weight. All fish with supplemental feeding appeared healthy. It is concluded that cow blood meal at 40% or 60% inclusion in diet can adequately replace fish-meal in S. niloticus supplemental diet in pond culture

An investigation of the effect of replacing fish meal with soyabean meal, groundnut cake and blood meal at varied proportion on growth and food utilization of the Clarias anguillaris fingerlings fed in outdoor hapas

Fingerlings of Clarias anguillaris obtained from a homogenous source through induce breeding and each with a mean weight of 2.8g were stocked in ten hapas each measuring 1.0x1.0m in outdoor concrete tank and were fed for eight (8) weeks. Results shows that the best growth rate was recorded among fingerlings fed fish meal as the only protein source (TD5) while DT2 containing soya bean, groundnut cake (40%), blood meal as the protein sources came next. The growth rate of fingerlings fed DT2 (40 % groundnut cake, 10% soyabean meal and 10% blood meal) was higher than those fed DT4 containing 10% fish meal but lower than those fed DT5 which has fish meal as its sole source of protein (53.0%). Analysis of various growth parameters like SGR, FCR and PER. shows that DT5 was the overall best diet but there was no significant statistical difference in weight gained by fish fed the five diets (P <0.05)

The effect of replacement of soyabean meal with blood meal on the growth of mudfish Clarias anguillaris (L) fingerlings

Blood meal and full fat soyabean meal were mixed in different proportions to give 0%, 10%, 25%, 50%, 75%, 90% and 100% meal in the protein fraction of the diet and fed to Claria anguillaris fingerlings in floating hapas. The growth performance of the fingerlings were monitored for 84 days. At the end of the experiment the mean weight of the fingerlings increased in the level of blood meal up to 50% blood meal in the diet after which there was a decline in the mean weight of the fish. This same level of blood meal gave the best specific growth rate, feed conversion efficiency and protein efficiency ratio. Thus the nutritive value of blood meal was enhanced by the addition of an equal level of full fat soyabean meal in the diet

The Circulation of Monterey Bay and Related Processes

(PDF contains 114 pages)

Physical circulation measurements in Pamlico Sound, used to locate new oyster sanctuaries

The population of eastern oyster, C. virginica, has declined over the last century on most areas of the east and gulf coasts. North Carolina’s restoration efforts depend on the construction of subtidal oyster reefs to be used as broodstock sanctuaries in Pamlico Sound, NC. Successful restoration of the oyster population requires several thriving reefs connected as a meta-population. C. virginica has a 2-3 week larval stage, during which it gradually settles through the water column. Larvae that can travel from one reef to another during that stage form the basis of a meta-population. (PDF contains 3 pages)

Flood circulation in oligomictic lakes: an undervalued reason for restriction of zooplankton production [Translation from: Memorie dell'Istituto Italiano di Idrobiologia Dott.Marco de Marchi 22 9-52, 1967]

Dormancy has been observed in several species of Cyclops. Animals in dormancy are characterised by almost complete inactivity of the main appendages and sluggish intestinal peristalsis. In Cyclops vicinus dormancy can take place in copepod stages III (19%), IV (78%) and V (3%) but one and the same animal only in one developmental stage. The author gives his own results between frequency of dormancy and photoperiod (20 C, 1000 lux). He concludes that dormancy in C. vicinus can be influenced by day length and can be ended prematurely by short day length.

Incidences de la modification de la circulation des eaux sur l'hydrochimie et le degré de contamination bacterienne d'un estuaire eutrophe tropical

In order to control the proliferation of floating aquatic vegetation in Côte d'Ivoire, a coastal inlet, allowing a direct communication between the Comoe river and the ocean, was created in September 1987. The impact of this operation on the hydrochemistry (salinity, nutrients, algal biomass) and the bacterial contamination level was studied in the area close to the Vridi canal.

La circulation superficielle dans la partie occidentale du Golfe de Guinée

Direct current measurements carried out for 10 years in the Gulf of Guinea clearly show the importance of transverse circulation. The general pattern of currents is characterized by an anticyclonic gyre. The seasonal variations of this circulation are calculated through the study of the 3-monthly averaged geostrophic currents. The biological consequences of such a circulation are outlined.

Circulation dans la partie orientale de l'Atlantique sud

The authors studied the circulation in the South Tropical Atlantic from hydrology and current measurements made during a cruise of the N.O. Capricorne in Nov 1971. Direct current measurements confirm the existence of the eastwards South Equatorial Counter-Current. The authors discuss the counter-current as confirmed by current measurements and high salinity core.