Differential gene expression of protein kinases in oocytes between natural gynogenetic silver crucian carp and amphimictic crucian carp

A modified mRNA differential display method has been applied to studying differential expression of protein kinase genes in oocytes between natural gynogenetic silver crucian carp and amphimictic crucian carp. Total RNA was reverse transcribed using downstream 3' primers T(12)MA, T(12)MG and T12MC respectively. Then the reverse transcription products were amplified using upstream 5' kinase-specific primer designed according to protein kinase conserved sequence. The PCR products had different patterns and numbers of: cDNA bands on polyacrylamide:gel. Totally 21 cDNAs fragments were recovered and cloned. Two of them were confirmed to be particularly expressed in oocytes of amphimictic crucian carp, and another was specific for gynogenetic silver crucian carp.

Gut contents of silver carp, Hypophthalmichthys molitrix, and the disruption of a centric diatom, Cyclotella, on passage through the esophagus and intestine

The digestibility of algae by stomachless filter-feeding fish has been debated for decades. Results from gut contents and digestive enzyme analysis suggest poor utilization, while the measurement of food assimilation using radiolabeled isotope techniques indicate reasonable assimilation efficiency. Phytoplankton in the gut contents of the planktivorous filter-feeding silver carp were studied during March-May. The fish were cultured in a large net cage in a shallow, hypertrophic subtropical Chinese lake, Lake Donghu. In terms of biomass, the dominant phytoplankton in the fore-gut contents were Cyclotella (average 77.8%, range 69.7-93.5%) and Cryptomonas (average 9.57%, range 0-20.4%). The Ivlev's electivity index E of silver carp was much higher for Cyclotella (1.54) than for Cryptomonas (0.56). The majority of the phytoplankton found in the intestines of silver carp were 8-20 mu m, but they were also able to collect particles as small as 4.5-10 mu m, smaller than their filtering net meshes, suggesting that the secretion of mucus may play an important role in collecting such small particles. We conclude that disruption of cell walls is principally by the pharyngeal teeth, explaining previous contradictory conclusions. (C) 1999 Elsevier Science B.V. All rights reserved.

Effect of natural dissolved humic material on bioavailability and acute toxicity of fenpropathrin to the grass carp, Ctenopharyngodon idellus

The effects of aquatic humic acids on the bioconcentration and acute toxicity of fenpropathrin were evaluated using grass carp, Ctenopharyngodan idellus, in laboratory freshwater systems. The results demonstrated that both bioavailability and acute toxicity decreased in the presence of aquatic humic acid 5 and 10 mg/liter. In addition, the extent of influence increased with increasing concentration of aquatic humic acid, (C) 1999 Academic Press.

Influence of dioxin and metal-contaminated sediment on phase I and II biotransformation enzymes in silver crucian carp

Several biochemical responses were measured in silver crucian carp (Carassius auratus gibelio) after exposure to sediments obtained from contaminated Ya-Er Lake, No, 1 pond, and an unpolluted reference site, Honglian Lake. After 1 week of exposure, a significant induction of the phase I biotransformation enzyme (ethoxylresorufin-o-deethylase, EROD) was found (83-fold of control), whereas the phase II biotransformation enzyme (glutathione S-transferase, GST) exhibited a slight, but significant induction (1,4-fold of control) after 4 weeks of exposure. The level of cellular glutathione in the liver was also slightly elevated after 4 weeks of exposure. The delayed response of GST to the contaminants indicates that the phase I and phase II biotransformation enzymes are regulated differently in fish. The results suggest that EROD is a sensitive bioindicator to assess the toxicity of dioxin-contamined sediment in the laboratory, (C) 1998 Academic Press.

Effects of silver carp density on zooplankton and water quality: Implications for eutrophic lakes in China

We examined the responses of zooplankton community, water transparency, chlorophyll a and nutrients to manipulation of density of silver carp (Hypophthyalmichthys molitrix) in an one-way factorial experiment using enclosures placed in Donghu (East Lake, 30 degrees 33' N, 114 degrees 23' E), located in Wuhan, P. R. China. Enclosures (18.75 m(3)) were treated with four silver carp densities, 0, 81, 225, 485 g/m(2). Total zooplankton abundance (excluding nauplii and rotifers except for Asplanchna sp.) and the mean size of dominant cladoceran species were significantly greater in enclosures with 0 and 81 fish densities than those in enclosures with 225 and 485 fish densities. Water transparency also improved significantly when silver carp densities were 0 or 81 g/m(2). We did not find significant effects of silver carp density on chlorophyll a, total phosphorus, or total nitrogen concentrations. We conclude that by reducing planktivorous fish to below the current density (190 g/m(2)), the zooplankton community can be shifted from the dominance of small-bodied Moina sp. to dominance of large-bodied Daphnia sp. Further, the water clarity can be increased.

A detection method for grass carp hemorrhagic virus (GCHV) based on a reverse transcription polymerase chain reaction

A rapid, sensitive and highly specific detection method for grass carp hemorrhagic virus (GCHV) based on a reverse transcription-polymerase chain reaction (RT-PCR) has been developed. Two pairs of PCR primers were synthesized according to the cloned cDNA sequences of the GCHV-861 strain. For each primer combination, only one specific major product was obtained when amplification was performed by using the genomic dsRNA of GCHV-861 strain. The lengths of their expected products were 320 and 223 bp, respectively. No products were obtained when nucleic acids other than GCHV-861 genomic RNA were used as RT-PCR templates. To assess the sensitivity of the method, dilutions of purified GCHV-861 dsRNA total genome (0.01 pg up to 1000 pg) were amplified and quantities of as little as 0.1 pg of purified dsRNA were detectable when the amplification product was analyzed by 1.5% agarose gel electrophoresis. This technique could detect GCHV-861 not only in infected cell culture fluids, but also in infected grass carp Ctenopharyngodon idellus and rare minnow Gobiocypris rarus with or without hemorrhagic symptoms. The results show that the RT-PCR amplification method is useful for the direct detection of GCHV.

EFFECT OF RATION SIZE ON THE GROWTH AND ENERGY BUDGET OF THE GRASS CARP, CTENOPHARYNGODON-IDELLA VAL

Young grass carp (12-13 g) were kept at five ration levels ranging from starvation to ad libitum feeding at 30-degrees-C. They were fed duckweed. Food consumption, absorption efficiency and growth were determined directly, and metabolism and nitrogenous excretion calculated indirectly from energy and nitrogen budgets, respectively. The relationship between specific growth rate and ration size was linear. Absorption efficiency for energy was not affected by ration size and averaged 50.6 +/- 0.57% (mean +/- s.e.). Depending on ration size, energy lost in excretion accounted for 4.5-5.9% of the food energy, energy channelled to metabolism accounted for 34.4-48.3% of the food energy, and energy retained as growth accounted for 6.7-11.9% of the food energy. Regardless of ration, a constant proportion of food energy (30.7%) was accounted for by feeding metabolism (total metabolism minus fasting metabolism). The energy budget at the maximum ration was: 100 C = 49.1F + 4.5U + 3.6R(fa) + 30.9R(fe) + 11.9G, where C, F, U, R(fa), R(fe) and G represent food consumption, faecal production, excretion, fasting metabolism, feeding metabolism and growth, respectively.

LABORATORY OBSERVATIONS ON THE CIRCADIAN FEEDING PATTERNS IN THE GRASS CARP (CTENOPHARYNGODON-IDELLA VAL) FED 3 DIFFERENT DIETS

Feeding intensities (number of bites per minute) were recorded each hour over a 24-h diel cycle for young grass carp fed three diets. The grass carp did not show distinct meals. Grass carp receiving plant diets (duckweed or elodea) fed almost continuously throughout the 24 h, while fish fed the animal diet (tubificids) ceased feeding or had very low feeding intensities for about a quarter of the diel cycle. The average feeding intensity in fish fed duckweed was three times higher than that in fish fed elodea and tubificids. Average dry matter intake per bite was much higher in fish fed the animal diet than in those fed the plant diets. In most individuals, there was no significant difference in feeding intensity between daytime and nighttime.

STUDY ON THE MODE OF PRIMARY CONTROL IN THE EGG OF GYNOGENETIC CRUCIAN CARP FOR INHIBITING THE DEVELOPMENT OF 2 TYPES OF SPERM NUCLEI

By comparing the different developmental characteristics of two types of sperm nuclei which were from gynogenetic fish (crucian carp) and amphimictic fishes (red carp, red goldfish and sex-reversal red carp) respectively in the eggs of gynogenetic crucian carp, it was preliminarily revealed that there existed selective inhibiting actions of the primary control in the eggs of crucian carp for inhibiting the development of the two types of sperm nuclei. To homologous sperms, the primary control showed weak effect, thus leading to the decondensation of homologous sperm nuclei at different degrees in the eggs of crucian carp. But to heterologous sperms, the primary control showed strong effects, resulting in the total inhibition of the development of heterologous sperm nuclei. Moreover, our experimental results also showed that the different developmental behavior of the two types of sperm nuclei might have a great relationship to the changes of the sex ratio in the population of gynogenetic crucian carp. The infiltration of "the genetic materials in sperm nuclei" into the female nucleus at random might play an important role in male emergence in the naturally gynogenetic population of crucian carp.