Hepatic Histopathological Characteristics and Antioxidant Response of Phytoplanktivorous Silver Carp Intraperitoneally Injected with Extracted Microcystins

Objective To investigate the hispathological characteristics and antioxidant responses in liver of silver carp after intraperitoneal administration of microcystins (MCs) for further understanding hepatic intoxication and antioxidation mechanism in fish. Methods Phytoplanktivorous silver carp was injected intraperitoneally (i.p.) with extracted hepatotoxic microcystins (mainly MC-RR and -LR) at a dose of 1000 mu g MC-LReq./kg body weight, and liver histopathological changes and antioxidant responses were studied at 1, 3, 12, 24, and 48 h, respectively, after injection. Results The damage to liver structure and the activities of hepatic antioxidant enzymes including catalase (CAT), superoxide dismutase (SOD), and glutathione peroxide (GPX) were increased in a time-dependent manner. Conclusion In terms of clinical and histological signs of intoxication and LD50 (i.p.) dose of MC-LR, silver carp appears rather resistant to MCs exposure than other fishes. Also, the significantly increased SOD activity in the liver of silver carp suggests a higher degree of response to MCs exposure than CAT and GPX.

Sequential ultrastructural and biochemical changes induced in vivo by the hepatotoxic microcystins in liver of the phytoplanktivorous silver carp Hypophthalmichthys molitrix

Up to now, in vivo studies on the toxic effects of microcystins (MCs) on the ultrastructures of fish liver have been very limited. The phytoplanktivorous silver carp was injected i.p. with extracted hepatotoxic microcystins (mainly MC-RR and -LR) at a dose of 1000 mu g MC-LReq. kg(-1) body weight, showing a time-dependent ultrastructural change in liver as well as significant increases in enzyme activity of plasma alanine aminotransferase (ALT), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH). We observed for the first time the occurrence of a large amount of activated secondary lysosomes, which might be an adaptive mechanism to eliminate or lessen cell damage caused by MCs through lysosome activation. Quantitative and qualitative determinations of MCs in the liver were conducted by HPLC and LC-MS2, respectively. MCs concentration in the liver reached the maximum (114.20 mu g g(-1) dry weight) after 3 h post-injection, and then rapidly dropped to 7.57 mu g g(-1) dry weight at 48 h, indicating a deputation of 99% accumulated MC-LReq. On the other hand, a decrease trend in glutathione (GSH) concentration was observed in the liver of silver carp while the activity of glutathione S-transferase (GST) increased significantly after injection. The high tolerance of silver carp to MCs might be due to the high basic GSH level in their liver, and/or an increased GSH synthesis. (C) 2007 Elsevier Inc. All rights reserved.

Biochemical and ultrastructural changes of the liver and kidney of the phytoplanktivorous silver carp feeding naturally on toxic Microcystis blooms in Taihu Lake, China

Many experimental studies have documented the impact of microcystins (MC) on fish based on either intraperitoneal injection, or oral gavaging via the diet, but few experiments were conducted by MC exposure through natural food uptake in lakes. In this study, the phytoplanktivorous silver carp were stocked in a large pen set in Meiliang Bay of Taihu Lake where toxic Microcystis blooms occurred in the warm seasons. Fish samples were collected monthly and MC concentrations in liver and kidney of the fish were determined by LC-MS. The maximum MC concentrations in liver and kidney were present in July when damages in ultrastructures of the liver and kidney were revealed by electron microscope. In comparison with previous studies on common carp, silver carp showed less damage and presence of lysosome proliferation in liver and kidney. Silver carp might eliminate or lessen cell damage caused by MC through lysosome activation. Recovery in the ultrastructures of liver and kidney after Microcystis blooms was companied with a significant decrease or even disappearance of MC. Catalase and glutathione S-transferase in liver and kidney of silver carp during Microcystis blooms were significantly higher than before and after Microcystis blooms. The high glutathione pool in liver and kidney of silver carp suggests their high resistance to MC exposure. The efficient antioxidant defence may be an important mechanism of phytoplanktivorous fish like silver carp to counteract toxic Microcystis blooms. (C) 2007 Published by Elsevier Ltd.

In situ studies on the bioaccumulation of microcystins in the phytoplanktivorous silver carp (Hypophthalmichthys molitrix) stocked in Lake Taihu with dense toxic Microcystis blooms

The phytoplanktivorous silver carp is an important biomanipulation fish to control cyanobacterial blooms and is also a food fish with the greatest production in China. The accumulation of the hepatotoxic microcystins (MCs) determined by LC-MS in various organs of silver carp was studied monthly in Lake Taihu dominated by toxic Microcystis aeruginosa. Average recoveries of spiked fish samples were 78% for MC-RR and 81% for MC-LR. The highest content of MCs was found in the intestine (97.48 mu g g(-1) DW), followed by liver (6.84 mu g g(-1) DW), kidney (4.8 8 mu g g(-1) DW) and blood (1.54 mu g g(-1) DW), and the annual mean MC content was in the order of intestine > liver > kidney > blood > muscle > spleen > gallbladder > gill. Silver carp could effectively ingest toxic Microcystis cells (up to 84.4% of total phytoplankton in gut contents), but showed fast growth (from 141 g to 1759 g in I year in mean weight). Silver carp accumulated less microcystins in liver than other animals in the same site or other fish from different water bodies at similar level of toxin ingestion. There was possible inhibition of the transportation of the most toxic MC-LR across the gutwall. Muscle of silver carp in Lake Taihu should not be consumed during period of dense Microcystis blooms while viscera were risky for consumption in more months. (c) 2006 Elsevier B.V. All rights reserved.

Effects of the phytoplanktivorous silver carp (Hypophthalmichthys molitrixon) on plankton and the hepatotoxic microcystins in an enclosure experiment in a eutrophic lake, Lake Shichahal in Beijing

The responses of nutrients, water transparency, zooplankton, phytoplankton and microcystins to a gradient of silver carp biomass (0, 18, 55, 110 g/m(3)) were assessed using enclosures in Lake Shichahai (Beijing). Picophytoplankton biomass increased with increasing fish stocking density (r=0.64, p=0.09). Silver carp significantly depressed zooplankton biomass, and thus, zooplankton grazing was too low to control phytoplankton. Intracellular microcystin (MC) content in the enclosures was correlated only to Microcystis biomass in the present study. Microcystis spp. biomass and intracellular microcystins content were much higher in lake water than those of enclosures with and without stocking fish. Stocking of silver carp could be an appropriate in highly productive Lake Shichahai, which naturally lacks of large cladoceran zooplankton. A fish stocking density of 55 g/m(3) was most efficient at controlling Microcystis blooms and increasing water clarity. Mean extracellular MC concentration in the lake water was almost the same with that of the enclosures with fish. (c) 2006 Elsevier B.V. All rights reserved.

Dynamics of microcystins-LR and -RR in the phytoplanktivorous silver carp in a sub-chronic toxicity experiment

A sub-chronic toxicity experiment was conducted to examine tissue distribution and depuration of two microcystins (microcystin-LR and microcystin -RR) in the phytoplanktivorous filter-feeding silver carp during a course of 80 days. Two large tanks (A, B) were used, and in Tank A, the fish were fed naturally with fresh Microcystis viridis cells (collected from a eutrophic pond) throughout the experiment, while in Tank B, the food of the fish were M. viridis cells for the first 40 days and then changed to artificial carp feed. High Performance Liquid Chromatography (HPLC) was used to measure MC-LR and MC-RR in the M. viridis cells, the seston, and the intestine, blood, liver and muscle tissue of silver carp at an interval of 20 days. MC-RR and MC-LR in the collected Microcystis cells varied between 268-580 and 110-292 mug g(-1) DW, respectively. In Tank A, MC-RR and MC-LR varied between 41.5-99.5 and 6.9-15.8 mug g(-1) DW in the seston, respectively. The maximum MC-RR in the blood, liver and muscle of the fish was 49.7, 17.8 and 1.77 mug g(-1) DW, respectively. No MC-LR was detectable in the muscle and blood samples of the silver carp in spite of the abundant presence of this toxin in the intestines (for the liver, there was only one case when a relatively minor quantity was detected). These findings contrast with previous experimental results on rainbow trout. Perhaps silver carp has a mechanism to degrade MC-LR actively and to inhibit MC-LR transportation across the intestines. The depuration of MC-RR concentrations occurred slowly than uptakes in blood, liver and muscle, and the depuration rate was in the order of blood > liver > muscle. The grazing ability of silver carp on toxic cyanobacteria suggests an applicability of using phytoplanktivorous fish to counteract cyanotoxin contamination in eutrophic waters. (C) 2003 Elsevier Ltd. All rights reserved.

Plasma biochemical responses of the planktivorous filter-feeding silver carp (Hypophthalmichthys molitrix) and bighead carp (Aristichthys nobilis) to prolonged toxic cyanobacterial blooms in natural waters

The planktivorous filter-feeding silver carp (Hypophthalmichthys molitrix) and bighead carp (Aristichthys nobilis) are the attractive candidates for bio-control of plankton communities to eliminate odorous populations of cyanobacteria. However, few studies focused on the health of such fishes in natural water body with vigorous toxic blooms. Blood parameters are useful and sensitive for diagnosis of diseases and monitoring of the physiological status of fish exposed to toxicants. To evaluate the impact of toxic cyanobacterial blooms on the planktivorous fish, 12 serum chemistry variables were investigated in silver carp and bighead carp for 9 months, in a large net cage in Meiliang Bay, a hypereutrophic region of Lake Taihu. The results confirmed adverse effects of cyanobacterial blooms on two phytoplanktivorous fish, which mainly characterized with potential toxicogenomic effects and metabolism disorders in liver, and kidney dysfunction. In addition, cholestasis was intensively implied by distinct elevation of all four related biomarkers (ALP, GGT, DBIL, TBIL) in bighead carp. The combination of LDH, AST activities and DBIL, URIC contents for silver carp, and the combination of ALT. ALP activities and TBIL, DBIL. URIC concentrations for bighead carps were found to most strongly indicate toxic effects from cyanobacterial blooms in such fishes by a multivariate discriminant analysis. (C) 2009 Elsevier B.V. All rights reserved.

Preliminary results of a silver carp breeding program in Vietnam

The article describes the silver carp (Hypophthalmichthys harmandi and Hypophthalmichthys molitrix) breeding program in Vietnam.

Mass breeding of silver carp in a cement cistern with crude H.C.G.

The paper presents a resume on experiments carried on mass breeding of the silver carp Hypophthalmichthys molitrix in a cement cistern through administration of crude human chorionic gonadotropin. The experiments proves that human chorionic gonadotropin (H.C.G.) can be used for mass breeding of the fish in simple enclosures, with facilities of some input and output of water and a dose of 5 mg/kg and 18 mg/kg of body weight, for males and females respectively are required for the purpose. Observations were made by using pituitary glands in similar way. For successful breeding of about 80 kg female together, the expenditure towards cost of pituitary gland and H.C.G. are respectively 275 and 117 Rs.

Polyculture of gulsha (Mystus cavasius Ham.) with rajpunti (Puntius gonionotus Bleeker) and silver carp (Hypophthalmichthys molitrix Val.) in earthen ponds

Culture of gulsha (Mystus cavasius) with rajpunti (Puntius gonionotus) and silver carp ( Hypophthalmichthys molitrix) was undertaken to assess the growth and production potential of these species under polyculture system. Fingerlings of gulsha, rajpunti and silver carp were stocked at a density of 18.000, 10.000 and 4.000/ha respectively. Two treatments were tested in this experiment. Treatment-I was conducted with rice bran and mustard oil cake and treatment-II with rice bran and duck weed. All the ponds were fertilized with urea and TSP at fortnightly intervals. After six months' rearing, the gross production was estimated to be 3,582 and 3,125 kg/ha from treatment-I and treatment-II respectively. Total yield showed non-significant differences (P>0.05) between the treatments.

Ice storage characteristics of cultured silver carp

Cultured silver carp (Hypopthalmichthys molitrix 800-1000 g) was stored in ice (fish to ice ratio 1:1) in a plywood box insulated with one inch thick expanded polystyrene and subjected to detailed examination of quality by chemical, microbiological and organoleptic evaluation at regular intervals to assess the storage life in good acceptable form. Alpha-amino nitrogen, non-protein nitrogen and pH values showed no positive correlation as spoilage index. Total volatile base nitrogen was not high at the end of the storage period although the fish became unacceptable during the period. There was steep decrease in total bacterial count during initial stages of storage and then increased steadily on further storage. Organoleptic evaluation of raw and cooked meat revealed that fish was in good acceptable form up to 14 days in ice.

Production and economic return in pangasiid catfish (Pangasius hypophthalmus) monoculture and polyculture with silver carp (Hypophthalmichthys molitrix) in farmers' ponds

The production of fish and net economic return in pangasiid catfish (Pangasius hypophthalmus) monoculture and polyculture with silver carp (Hypophthalmichthys molitrix) in farmers' ponds were assessed. The experiment was arranged in three treatments each with three replications. The ponds were stocked with 30,000 fishes per hectare. In treatment 1 (T1) pangasiid catfish only, in treatment 2 (T2) pangasiid catfish and silver carp at the ratio of 1:1, and in treatment 3 (T3) pangasiid catfish and silver carp at the ratio of 2:1 were stocked. At harvest, production of fish was found significantly (p<0.05) different among the treatments, highest in T1 and lowest in T2. Though the total biomass production and total economic return was significantly highest in T1 than in T2 and T3, the net economic return was lowest because of the required highest input costs especially for supplemental feed and fingerlings, resulted the highest cost per unit yield (CPY in Tk/kg) in T1. Highest cost for supplemental feed required in T1 was due to highest quantity of feed required for the highest number of pangasiid catfish stocked in that treatment. The findings of the present study suggest that though monoculture of pangasiid catfish give higher fish biomass production but polyculture with silver carp is environmentally good and economically profitable.

Marketing of value-added products from silver carp and involvement of rural people in the production and marketing chain

Cost-profit analysis and market testing of some value-added products from silver carp such as fish mince block, fish sausage, fish ball, fish stick and fish burger were analyzed during April 2001 to March 2002. The study also explored the possibility to involve rural low-income people in the production and marketing of such products. The production of silver carp was higher in greater Jessore and Mymensingh districts but the price remained low during the peak-harvesting season in October to November. The price varied with size of the fish, season, market characteristics and effective demand of the buyers. Price of about 500 g size fish was found to be Tk. 20-25/kg in the rural markets. The average size of fish in the rural markets was 3S0-550 g while that in the urban markets it was 700-1,200 g. The cost of production of the value added products and profit margin were assessed on the basis of market price of the raw material as well as that of the finished products, transportation, storage and marketing costs. The profit margins of 34%, 39%, 81% and 31% of their sales price were obtained for fish sausage, fish ball, fish stick and fish burger, respectively. Actual production cost could be minimized if the fish is purchased directly from the farmers. Consumer's acceptance and marketability tests showed that both rural and urban people preferred fish ball than fish sausage. However, response towards the taste, flavor and color of fish ball and fish sausage was found to vary with occupations and age of the consumers. A correlation was observed between age group and acceptance of new products. Fish ball, fish stick and fish burger were found to be the most preferable items to the farmers because of easy formulation process with common utensils. Good marketing linkage and requirement of capital had been identified as the prerequisites for operating small-scale business on value-added fish products.

Studies on the suitability of producing value-added products from silver carp, Hypophthalmichthys molitrix

Silver carp, Hypophthalmichthys molitrix is contributing significantly to the total production of fish through aquaculture in Bangladesh. However, its low market price has become a serious concern to the fish farmers. The suitability of silver carp mince for the production of various value-added products (VAPs) - surimi, fish sausage, fish burger and fish stick was studied during April-September 2000 to ensure more appropriate and profitable utilization of silver carp. Surimi/frozen mince block was produced by washing the silver carp mince with 0.1% NaCl for 7-8 min (4-5 min agitation and 3-4 min settling). A two-step heating schedule for incubation at 50°C for 2 h and cooking at 95°C for 30 min gave high textured good quality consumer product. With the addition of cryoprotectants, surimi could be kept frozen for 5 months without loosing [sic] much of its textural and sensory qualities. Mince-mix and a batter with different ingredients and spices were formulated to produce fish burger using potato smash as the binding agent. Fish flake-mix and a batter with different ingredients and spices were formulated to prepare fish stick using both potato starch and potato smash as filler ingredients. Unwashed and washed frozen mince block or fresh flesh of silver carp was used to prepare fish sausage by heating at 100°C for 1 h after incubating at 50°C for 2 h. A spice-mix formulated with various local spices at the rate of 1.0-1.2% gave good texture and flavor to the sausage. A good-appeared sausage-pink color was developed by combining three food-grade colors of asthaxanthin. Products prepared with potato starch, potato smash and rice smash had an acceptable bacterial load in refrigeration (5°C) for up to 8 days and in room temperature (28°C) for up to 3 days. No coliform bacteria were found in the products prepared.

Study on shelf life of fish cake prepared from surimi of silver carp (Hypophthalmichthys molitrix) during frozen storage

Surimi was prepared from silver carp with an aim to put this underutilized fish for profitable use. The mince prepared was washed twice with chilled water (5°C) using mince to water ratio (w/v) of 1:2 for 5-6 minutes each. After final dewatering to moisture content to about 80%; half the quantity of washed minced meat was mixed with cryoprotectants (4% sorbitol, 4% sucrose and 0.3% sodium tripolyphosphate) to produce surimi. The prepared surimi and the dewatered minced meat were packed in LDPE bags, frozen using a plate freezer and stored at -20°C. Surimi and dewatered minced meat from frozen storage were used as base material for production of fish cakes. These were fried at 160°C for 3 to 4 minutes before serving for organoleptic test. Changes in salt soluble nitrogen, total volatile base nitrogen, non-protein nitrogen, peroxide value and free fatty acid of surimi and dewatered mince were estimated at every ten days interval during the storage period of 3 months. The study has indicated that frozen storage of surimi could be a potential method for effective utilization of silver carp. This surimi when incorporated in fish cakes yielded products which retained the shelf life even up to 90 days of storage.