Determination of threshold current densities for different reactions of fishes using a pantostat

The threshold current densities required for first reaction, galvanotaxis and galvanonarcosis of fish depended upon species, length of the body, conductivity of water, nature of current and frequency of impulses. The threshold values and their ratios decreased with increase in length of fish. With rise in conductivity of water in the ratio of 1:4:13, these values increased in the ratio 1: 2:5. Impulse D. C was superior to continuous D. C and the threshold values of current densities for different reactions of fish decreased with rise in impulse frequency reaching minimum at an impulse frequency of 48/sec. Among Salmo irideus, ldus melanotus and Cyprinus carpio, the first one was affected earlier and required minimum current densities to exhibit the reactions, while the last one showed similar reactions only at higher current densities.

On the reaction of marine fishes in interrupted A.C. of 50 Hz

The reactions of marine fishes in interrupted A. C. off uniform electrical field and constant current density have been studied in order to determine the optimum effective periods of shock: for narcosis and fixation. Higher effective periods were required with gradual decrease in potential difference between head and tail, which in turn depended on the length of fish and its position in electrical field. The body voltage of fish varied directly with the length and inversely with the rise of angle between their body axis and field lines. The fish subjected to higher effective period took longer time for recovery from narcosis. The effect of impulse current was manifested by increased rate of gill movements in Platax tefora, Haetodon ollaris, Crysophus burda, Siganus vermiculatus and Scolopsis leucotaenia, and lowered rate of movements in others when subjected to different effective periods inspective of length.

Reaction of fishes to the underwater A.C. field

A comparative study on the effect of A.C. field on Puntius ticto, Heteropneustis fossilis and Tilapta mossambica was carried out using a slowly rising field intensity. Well defined reactions appeared in the species of fish with slight specific variations, depending on their orientation in the electrical field, on reaching the field intensity to specific value. These reactions can be distinguished as first reaction, when the fish perceive the surrounding field, jerky swimming when parallel to the current lines (longitudinal oscillotaxis), the static position finally adopted by the fish sooner or latter depending on the potential gradient (transverse oscillotaxis), and a state of muscular rigidity (tetanus). After switching off the current, a hypnotic condition prevailed in the treated fishes before returning to their normal swimming condition. The orientation of fish body in the field had an important bearing on the behaviour reactions and current thresholds necessary for those reactions. Initial reaction, jerky swimming between electrodes and hypnosis after stoppage of current appeared in fishes earlier when the fish body was in parallel to the current lines, whereas fishes responded to transverse oscillotaxis quickly when perpendicular to current lines.

Report on the occurrence of one subspecies of Scylla serrata (Forskal) in Cochin backwaters

The taxonomy of the mud crab, Scylla serrata has become a topic of interest. Alcock (1899) described this species from Indian waters. It was considered that under the genus Scylla, only one species, namely, Scylla serrata was valid. But Estampador (1949) revised the genus and reported the occurrence of four sorts, Scylla serrata, S. tranquebarica, S. oceanica and a variety S. serrata, var paramarnosain in Philippine waters. He established his claim based on the morphology such as colouration, relative length of chelae and also on the cytological events during gametogenesis. Serene (1952) pointed out that two distinct categories under this genus were demonstrable on the basis of colouration. But Stephenson & Campbell (1960) concluded them as only synonyms, but pointed out the need for further investigations.

Merging effects of vitamin E and C on biological factors, blood characteristics and resistance against thermal stress in rainbow trout (Onchorhyncus mykiss)

This study was conducted to assay the effects of different levels of dietary vitamins C and E on growth indices and survival and resistance against thermal stress of rainbow trout (Oncorhynchus mykiss) in pond culture of Marzan abad from December 2011 to February 2011. Seven diets were supplemented. 300 fish with the average weight of 17 g were introduced to ponds for 60 days. The results showed that the highest and the lowest weight gain were in fish fed with diet containing 50 mg/kg vitamin C and E and 0 mg/kg vitamin C and E(control) , respectively. The highest and the lowest Feed Conversion Ratio (FCR) were measured in control and diet 50 mg/kg vitamin C and E. There is a significant difference in their treatments (P<0.05). Also, the lowest and highest amount of Weight Gain (WG) were observed in (E) treatment with 165.04% and 117.5% in control, the highest and lowest Specific Growth Rate (SGR), Protein Efficiency Ratio (PER), Condition Factor (CF) was found in control and treatment 50 mg/kg vitamin C and E, respectively(P<0.05). In conclusion vitamin C and E have an important role in enhancement of growth performance and feed efficiency of rainbow trout.The highest red blood cells were found in combined treatments and which the vitamin C was added.The highest RBC were found in E treatment(1.1×104 /mm3) and the lowest one in control (P˂0.05). Counting white blood cells also confirmed highest quantity in combined treatments with (69.83×104/mm3) and the lowest one (28.83×104 /mm3) in control. In conclusion these vitamins have a significant role in blood characteristics. Meantime, the resistance against termal stress was measured at the end of 60 days by facing fishes into 5 centigrade warmer water so consentration of Cortisol and Glucose measured for this reason.The lowest cortisol amount was measured in E treatment with 188.74 ng/ml and the highest was found in control(P<0.05). There was a significant difference in blood glucose consentration of fishes in F treatment with (78.66 mg/dl) and control with 136 mg/dl as a highest one(P<0.05).

Histology and hormonal effect of Ghrelin on ovary of Benni (Barbus sharpeyi) in order to study sexual maturity, zygote and larval generation

Benni (Barbus sharpeyi) is valuable fish that Khuzastan fisheries office propagated it artificially in Susangerd Fish Propagation Center every year. Pituitary gland is used for this aim but female fish lost their fertilization power after 2-3 years, so in present research, new hormone, that is called Ghrelin. The aims of this research are histology, hormonal, zygote and larval generation studies and comparing the results with each other. Ghrelin is a multifunctional peptidyl hormone which increases GTH-II in fish, amphibian, and birds and mammalian so its effect on Benni sexual maturation was studied. Human Ghrelin (hGRL) was obtained from ANASPEC, Canada, with 28 amino acids. In the present study, three levels of ghrelin including 0 (sham treatments), 0.10 (treatment 1) and 0.15 μg/g (treatment 2) body wt and one level of pituitary gland 4000 μg/g (pituitary treatment) with two replications were used. 56 specimens were injected intraperitonealy and their ghrelin level was evaluated immediately after injection and after 24 h. Control fish(n=16) were just injected by physiological saline. For hormonal studies sham and experimental fish(n=40) were anesthetized with MS-222 at a concentration of 250 mg l-1, and blood samples were collected and kept at 4ْC, then spun to collect serum. Serum samples were stores at -20ْC until the RIA for CTH-II. For histology studies immediately after injection a piece of ovary was collected from control fish (Sham zero) after being anesthetized. The sampled ovaries were fixed in Buin solution and embedded in paraffin, and stained to Sections of 5–6 μm using haematoxylin and eosin. The ovarian samples were performed with a compound microscope. Histology and micrometry studies had done. The mature oocytes had given from mature fish, then weighted and the working fecundity were counted. The mature oocytes fertilized, the eggs were incubated and the percentage of fertilization was calculated. After 72h the eggs hatched and the percentage of hatch was counted. The percentage of hindrance was calculated after 6 days. Hormonal results indicate that ghrelin and pituitary increase significantly the GTH-II level in comparison to sham. Macroscopic observations (before taking ovary) showed that ovaries with green colored have couple oval structure located in the abdominal cavity. Microscopic studies of dissected ovaries indicated simultaneous growth of 127 oocytes with 6 stages. The type of the ovary is asynchronous. The results indicated that both of the ghrelin treatment increased the percentage of mature follicles followed by decrease of immature follicles. There were significant differences (P<0.05) between the number of mature and immature follicles. Average diameter of follicle in both of the ghrelin treatment was significantly (P<0.05) declined in the stages of the vitellogenesis when the result compared to the other treatment. Just treatment 1 and pituitary treatment can give mature oocytes. The fecundity of pituitary treatment significantly increase in comparision to ghrelin treatment (P<0.05). In food-restricted fish where endogenous ghrelin levels are known to be increased, a chronic administration of ghrelin induces overt negative effect in releasing mature oocytes. The percentage of fertilization was significantly increase (P<0.05) in ghrelin t. in comparison to pituitary t. and the percentage of hatch was significantly increase (P<0.05) in pituitary t. in comparison to ghrelin t. There was no significant difference (P>0.05) in terms of percentage of hindrance between treatments. In conclusion, the present study demonstrated that ghrelin has positive effect on the level of GTH-II, oocyte maturation, ovarian vitellogenesis and the number of mature follicles of Barbus sharpeyi ovary. Increasing of the mature follicles number reduces their average diameter, indicating stimulating effect of ghrelin in sexual maturation of Barbus sharpeyi.The ghrelin and pituitary treatment have equal chance in the post-stage of spawning.

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 .

Survey of biosystematic and sexual reproductive physiology in Holothuroidea on Hormozgan Province

Sea cucumbers belong to phylum Echinodermata, order Holothuroidea are an abundant and diverse group of Invertebrates, with over 1400 species occuring from the intertidal to the deepest oceanic trenches. Sea cucumbers are important components of the food chain in temperate and coral reef ecosystems and they play an important role as deposite feeders and suspension feeders. Rapid decline in populations may have serious consequences for the survival of other species that are part of the same complex food web,as the eggs, larve and juveniles constitute an important food source for the other marine species including crustaceans, fish and mollusks. In addition sea cucumbers are often called the earthworms of the sea, because they are responsible for the extensive shifting and mixing of the substrate, and recycling of detrital matter. Sea cucumbers consume and grind sediment and organic material into finer particles , turning over the top layers of sediment in lagoons , reefs and other habitats and allowing the penetration of oxygen. While the taxonomy of the holothurian families is generally well known , the distinction of similar species is difficult. There are relatively few holothurian taxonomist.Most sea cucumber species can be identified by Holothurin taxonomists by using the calcareous skeletal ossicles found in the body wall. In this study , at first a sea cucumber from Kish island in Persian gulf has recognized. Individuals collected from west and east extend far away into north and south of coral reefs by diving. I have checked them morphologically and anatomically.Then with key to the orders of the Holothuroidea, They belong to the Aspidochirotida with key to the families of Aspidochirotida, they were in Stichopodidae families and with key to the genus of Stchopodidae, they were Stichopus. Then ossicles were extracted at National Museum of Natural History, by Dr David Pawson. The ossicles were measured on a transect across a slide prepared from the mid-dorsal region of each specimen.The one we have in the shallow waters of Kish island, is Stichopus hermanni, a massive holothurian, body broad, considerably flattened ventraly ,the dorsal side slightly arched and the lateral sides almost vertical; body wall fairy thick and soft ; mouth subterminal; anus central; tentacles usually 20 in number of length and leaf shaped. Numerous ossicles consisting of table with large discs having usually 7 to 15 peripheral holes, but often irregular or incomplete and spire of moderate height ending in a group of spinelets, rosettes of variable development, and c-shaped rods. Color (exept papillae)partly remained after preservation in alcohol which is found at the depth of 4 to 8 meters, on coral reef. Furthermore, the sexual reproductive cycle was described using standard methods. Gonads were removed and transferred to Bouin's fixative for four weeks and then processed according to standard embedding technique. To prevent the loss of tubule contents during embedding, the tubule sections, were cut well beyond the segment selected for sectioning. For each individual, six sections, each section with 5µm diameter by microtome were cut from tubules. These sections were first placed on gelatin coated slides (the gelatin was heated to 42°c) and then transferred to the oven at 37°c for one hour. This technique usually prevents the fragil tubules from breaking and the loss of gametes. The slides were stained with Eosin and Hematoxylin, and good resolution of the various cell types achieved.A second series of slides was stained with the Periodic Acid Schiff(PAS) to identify polysaccharides(glycogen). Monthly sampling was occurred.The sexual reproductive cycle was defined through the combined use of these criteria: Monthly percentages of the gonad stages for each sex, the monthly gonad index (GI) , given as the ratio of the wet gonad weight (G) to the dray weight (DW)and the monthly percentage of individuals that undetermined sex. The gonad consists of two tufts of tubules on which saccules develop. Gonadal development was classified into five stages: post spawning, recovery, growth, advanced growth, and mature stage that were adapted from the earlier studies of holothurians. Histological preparations showed that the sex of larger individuals could be identified by the presence of oogonia and young oocytes in females, and spermatogonic stages in males.The mean diameter of the tubules and gonadal mass follow annual cycles, increasing from late winter through spring, and dropping abruptly after spawning in the summer. Gametogenesis is generally a prolongate process and begins in March. By summer the ovarian tubules contain oocytes with diameter of 120-240 pm and the testicular tubules contain an abundance of spermatozoa (diameter 5-6 gm ).Following spawning the predominant activity within the spent tubules is phagocytosis of the residual gamets.The active phase of gametogenesis (March to July), coincides with an increasing photoperiod regim, and an accelerated gametogenesis occurs in July when temperature is high. Throughout the year, the gonad of Stichopus hermanni is larger in males than in females, and this is due to the number of tubules in the testis rather than to tubules length or diameter.