Electrochemical measurement of hydrogen peroxide in plasma: Evaluation of freshwater prawn (Macrobrachium rosenbergii) and crucian carp (Cyprinus carpio) phagocytes under natural conditions

An electrochemical technique for the real-time detection of hydrogen peroxide (H2O2) was employed to describe respiratory burst activity (RBA) of phagocytes in plasma which can be used to evaluate the ability of immune system and disease resistance. The method is based upon the electric current changes, by redox reaction on platinum electrode of extracellular hydrogen peroxide (H2O2) released from phagocytes stimulated by the zymosan at 680 mV direct current (d.c.). Compared with the control, activation of respiratory burst by zymosan particles results in a high amperometric response, and a current peak was obtained during the whole monitoring process. The peak current was proved by addition Of Cu2+ and other controls, to be the result of intense release of H2O2 from phagocytes. The peak area was calculated and used to evaluate the quantity of effective H2O2, which represents the quantity of H2O2 beyond the clearance of related enzymes in plasma. According to Faraday's law, the phagocytes' ability of prawns to generate effective H2O2 was evaluated from 1.253 x 10(-14) mol/cell to 6.146 x 10(-14) mol/cell, and carp from 1.689 x 10(-15) Mol/Cell to 7.873 x 10(-1)5 mol/cell. This method is an acute and quick detection of extracellular effective H2O2 in plasma and reflects the capacity of phagocytes under natural conditions, which could be applied for selecting species and parents with high immunity for breeding in aquaculture. (c) 2007 Elsevier Ltd. All rights reserved.

Thermoaclimatory variations in the microenvironment of hemoglobin in the rainbow trout, Salmo gairdneri and the carp, Cyprinus carpio

Several inorganic substances (e.g., C£ , Mg , Ca , H ) are potent negative modulators of hemoglobin-oxygen affinity. To evaluate the possibility that potentially adaptive changes in the red cell ionic environment of hemoglobin may take place during acclimation of fishes to increased environmental temperature, hematological status (hemoglobin, hematocrit, red cell numbers, mean erythrocytic volume and hemoglobin content), plasma + + 2+ 2+ and packed red cell electrolyte levels (Na , K , Ca , Mg , C£ ) were evaluated in summer and winter populations of the stenothermal rainbow trout, Salmo gairdneri, following acclimation to 2°, 10°, 18°C, and in a spring population of eurythermal carp, Cyprinus carpio, held at 2°, 16° and 30°C. From these data cell ion concentrations and ion:hemoglobin ratios were estimated. In view of the role of red cell carbonic anhydrase in the reductions of blood C02 tensions and the recruitment of Na and C£~ lost by fishes, a preliminary investigation of thermoacclimatory changes in the activity of this system in rainbow trout erythrocytes was conducted. Few changes in hematological status were encountered following acclimation. There was, however, some evidence of weight-specific differential hematological response in carp. This lead to markedly greater increases in hemoglobin, hematocrit and red cell numbers in smaller rather than in larger specimens at higher temperatures; variations which were 2+ well correlated with changes in plasma Ca . Plasma composition in summer trout was not altered by acclimation. In winter trout plasma Na and K increased at higher temperatures. Carp were characterized by increases in plasma calcium, and reductions in sodium and magnesium under these conditions. Several significant seasonal differences in plasma ion levels were observed in the trout. (n) In trout, only erythrocytic K and K :Hb were altered by acclimation, rising at higher temperatures. In carp Na , Na :Hb, C£~ and C£~:Hb in- 2+ 2+ creased with temperature, while Mg and Mg :Hb declined. Changes in overall ionic composition in carp red cells were consistent with increases in H content. In both species significant reciprocal variations in C£~ 2+ - + and Mg were found. In mammalian systems increases in C£ and H reduce hemoglobin-oxygen affinity by interaction with hemoglobin. Reduction in 2+ 2+ Mg maximizes organophosphate modulator availability by decreasing ATP»Mg complex formation. Thus, the changes observed may be of adaptive value in reducing hemoglobin-oxygen affinity, and facilitating oxygen release to cells at higher temperatures. Trout appear to maintain a high chloridelow magnesium state over the entire thermal tolerance zone. Carp, however, achieved this state only at higher temperatures. In both species mean erythrocytic volume was decreased at higher temperatures and this may facilitate branchial oxygen loading. Since mean erythrocytic volume was inversely related to red cell ion content, it is hypothesized that reductions in cell volume are achieved by export of some unidentified solute or solutes. Variations in the carbonic anhydrase activity that could be attributed to the thermoacclimatory process were quite modest. On the other hand, assays performed at the temperature of acclimation showed a large temperature effect where under in vivo conditions of temperature fish acclimated to higher temperatures might be expected to have higher activities. Furthermore, since hematocrit increased with temperature in these fish, while carbonic anhydrase is present only in the erythrocyte, the whole blood levels of this enzyme are expected to increase and further augment the temperature effect. This, in turn, could aid in the reduction of C02 (111) tension and increase the production of H and HC0~~ used in the active uptake of Na and C£ at higher temperatures.

Distribution, extracellular virulence factors and antibiogram of motile aeromonads in fresh water ornamental fishes and immune response of Cyprinus carpio against Aeromonas hydrophila infection

The thesis deals with the prevalence and distribution of motile aeromonads in selected ornamental fishes. The presence of motile aeromonads in ornamental fishes and associated carriage water is well documented. Though aeromonads are a part of autochthonous flora of natural waters, disease outbreak occurs as a result of environmental stress on the cultured species and virulence of the pathogens. While ornamental aquaculture in many parts of the world is highly organized and practiced scientifically, it is highly unorganized in India. The culture ponds/tanks are often maintained in very poor manner and the fishes are subjected to high degree of stress during transportation from the production facility to retail vendors. The situation is no better at retail outlets, where fishes are maintained in crowded condition without proper aeration or food. All these could result in high prevalence of diseases caused by motile aeromonads. No systematic study has been carried out to understand the prevalence of motile aeromonads in ornamental fishes and carriage water . It also gives an account of the production of extracellular virulence factors and the antibiogram of the different species of motile aeromonads isolated. The growth characteristics and virulence potential of a representative strain of Aeromonas hydrophila is also studied. The nucleotide sequencing of the strain was carried out and sequences deposited in Genbank. Survival and immune response of Cyprinus carpio under different stress conditions and on probiotic treatment with Bacillus NL110, when challenged with A. hydrophila is also dealt within this thesis.

A comparison of trace element concentrations in cultured and wild carp (Cyprinus carpio) of Lake Kasumigaura, Japan

The concentrations of 13 elements were determined in the muscle, liver, intestine, kidney, and gonads of cultured and wild carp caught at two sites in Lake Kasumigaura, Japan, between September 1994 and September 1995. Despite having a reputation for being heavily polluted, the carp were not heavily burdened with metals. Our results suggest that despite their dietary differences, the wild and cultured fish were accumulating and distributing metals in the same manner and that aquaculture practices are not increasing metal concentrations in these fish. Metal concentrations were lowest in muscle, and did not exceed established quality standards for fish. The differences in metal concentrations between cultivated and wild carp are negligible and should pose no health problems for consumers of either type of fish.

Using mitochondrial nucleotide sequences to investigate diversity and genealogical relationships within common carp (Cyprinus carpio L.)

Direct sequencing of mitochondrial DNA (mtDNA) D-loop (745 bp) and MTATPase6/MTATPase8 (857 bp) regions was used to investigate genetic variation within common carp and develop a global genealogy of common carp strains. The D-loop region was more variable than the MTATPase6/MTATPase8 region, but given the wide distribution of carp the overall levels of sequence divergence were low. Levels of haplotype diversity varied widely among countries with Chinese, Indonesian and Vietnamese carp showing the greatest diversity whereas Japanese Koi and European carp had undetectable nucleotide variation. A genealogical analysis supports a close relationship between Vietnamese, Koi and Chinese Color carp strains and to a lesser extent, European carp. Chinese and Indonesian carp strains were the most divergent, and their relationships do not support the evolution of independent Asian and European lineages and current taxonomic treatments.

Use of pineapple juice for elimination of egg stickiness of common carp (Cyprinus carpio L.)

Four treatments including three different concentrations of pineapple solution (I %, 3%, 5%) and the traditional method (salt/urea/tannin solutions) were used to eliminate egg stickiness of common carp. With the traditional method, fertilization and hatching rates were 74.5% ± 1.2 and 70.2% ± 1.1, respectively. The highest fertilization rate (89.3% ± 0.7) and hatching rate (86.6% ± 1.4) were found with treatment of 1% pineapple juice solution and was significantly different from those obtained with other treatments (P<0.05). Using pineapple juice for desticking common carp eggs also reduced the time of egg handling from almost one hour required with the traditional method, to about three minutes, as well as increasing fertilization and hatching rate under hatchery conditions.

The effect of seed morphology on the potential dispersal of aquatic macrophytes by the common carp (Cyprinus carpio)

1. The potential for seed dispersal by fish (ichthyochory) will vary among aquatic plants because of differences in seed size and morphology.

2. To examine how seed morphology influences the probability of dispersal by the common carp (Cyprinus carpio), we studied seed ingestion, retention time and subsequent egestion and germination of seeds of Sparganium emersum and Sagittaria sagittifolia, two aquatic plant species with similar sized but morphologically different seeds.

3. We compared dispersal probabilities between the two plant species, in which the probability of dispersal is assumed to be a function of the probabilities of seed ingestion, egestion and germination, and the dispersal distance is assumed to be a function of seed egestion rate over time.

4. We found that, although the soft seeds of S. sagittifolia had an approximately 1.5 times higher probability of being ingested by the carp than the hard seeds of S. emersum (83.15% ± 1.8% versus 56.16% ± 2.7%, respectively), the latter had an almost twofold higher probability of surviving the passage through the digestive tract (38.58% ± 2.7% versus 20.97% ± 1.5%, respectively). Patterns of seed egestion over time did not differ between the two plant species, despite the difference in seed morphology. Gut passage had a different effect on seed germination between plant species. Compared with non-ingested controls, seeds of S. emersum showed a 12.6% increase in germination and a 2.1 day acceleration in germination rate, whereas seeds of S. sagittifolia displayed a 47.3% decrease and 5.1 day delay, respectively.

5. Our results suggest that seed morphology affects the dispersal probability and postdispersal establishment, but not the dispersal distance, of aquatic plants that are dispersed by fish.

Germ line Cysts and the Formation of the Germinal Epithelium During the Female Gonadal Morphogenesis in Cyprinus Carpio (Teleostei: Ostariophysi: Cypriniformes)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Histological study of the dynamics in epidermis regeneration of the carp tail fin (Cyprinus carpio, Linnaeus, 1758)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

In vitro exposure to fullerene C60 influences redox state and lipid peroxidation in brain and gills from Cyprinus carpio (Cyprinidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Effects of acute heat exposure on respiratory metabolism and blood glucose in freshwater fishes, Prochilodus scrofa (curimbatá) and Cyprinus carpio (carp) acclimatized to tropical winter

1. 1. Routine oxygen consumption and blood glucose were determined from freshwater fishes, Prochilodus scrofa and Cyprinus carpio, exposed at high temperatures for 1 hr. 2. 2. Prochilodus scrofa had a significantly higher rate of oxygen consumption at 30°C than at 25°C, and carp higher at 25°C than at 30°C. 3. 3. Blood glucose was significantly higher for Cyprinus carpio than for Prochilodus scrofa at 25 and 30°C; however, after exposure to these temperatures for 1 hr blood glucose did not change significantly for both species. 4. 4. The results suggest that these interspecific variations may be linked to the differences between native and foreign fishes and their way of life. © 1985.

Células sangüíneas, eletrólitos séricos, relação hepato e esplenossomáca de carpa-comum, Cyprinus carpio (Cyprinidae) na primeira maturação gonadal

This research studied the condition (Kn) relative factor, the hepatosomatic (HSR), the splenosomatic (SSR) relation, the erythrocytes, thrombocytes and leukocytes number, the glicogen locality by PAS method in thrombocytes and leucocytes and the total protein and electrolytes serices levels in both Cyprinus carpio L. sex. The female Kn showed higher value than the males. The female HSR average values were lower than the ones observed in males, while the leukocytes, lymphocytes and neutrophils percentage number were the highest. However, the males magnesium and choride serices levels were higher than the females. The HRS average values, hematocrit, haemoglobin concentration, average corspucular volume (ACV), average corpuscular haemoglobin concentration (ACHC), erythocytes number, trombocytes, monocytes, oesinophils, positive PAS leukocyte granular (PAS-LG), and total protein, sodium, potassium and calcium serices levels did not show significant statistics difference (p> 0.05) between males and females. The PAS method showed glicogen granules into the thronbocytes, eosinophils and neutrophils cytoplasm. Nevertheless, in the PAS-LG cytoplasm, neutral glicoprotein granules were also observed. Monocytes and lynfocytes showed negative reaction to PAS. In the teleosts, the gonodas matureness presents higher relation with the factor of condition. During the reprodution period it is unlikely to find the sexual effects isolated in the hemogram, because most of the time that is not the only factor to cause dimorphism.

Estudo da ação da dexametasona na regeneração da nadadeira caudal do peixe teleósteo Cyprinus carpio (carpa)

Pós-graduação em Ciências Biológicas (Biologia Celular e Molecular) - IBRC

Nicho espermatogonial em Cyprinus carpio e papel do Amh na espermaogênese

Pós-graduação em Aquicultura - FCAV

A technique for the determination of the available oxygen in living carp (Cyprinus carpio) muscle

1. 1. A polarographic method for the measurement of the available oxygen in the muscle of living carp by the use of a platinum microelectrode is proposed. 2. 2. The oxygen and the reference electrodes were assembled in a single insertion piece which was implanted in the muscle of a living carp maintained in a special experimental chamber. 3. 3. Curves for normal oxygen levels corresponding to air-saturated water, as well as to a carbogene-saturated water, were obtained. 4. 4. The method can be considered adequate for the measurement of tissue oxygen in living fishes. © 1984.