Inductive transcription and protective role of fish heme oxygenase-1 under hypoxic stress

Heme oxygenase-1 is the rate-limiting enzyme in the degradation of heme into biliverdin, carbon monoxide and free divalent iron. In this study, we cloned heme oxygenase isoform 1 (CaHO-1) from a hypoxia-tolerant teleost fish Carassius auratus. The full-length cDNA of CaHO-1 is 1247 bp and encodes a protein of 272 amino acids. RT-PCR and real-time PCR analysis indicated that CaHO-1 was predominantly transcribed in posterior kidney, head kidney, gill and intestine, and induction of gene transcription was observed predominantly in posterior kidney under hypoxic stress. Moreover, the hypoxia-induced transcription was confirmed in goldfish larvae and in in vitro cultured CAB cells. Fluorescence of the HO-1-GFP fusion protein revealed a cytoplasmic and plasma membrane localization, which was consistent with the putative transmembrane structure. Subsequently, we established a stably transfected CAB/pcDNA3.1-HO-1 cell line and a control CAB/pcDNA3.1 cell line, and found that the number of dead cells was obviously reduced in the pcDNA3.1-HO-1-transfected group following 4 days of hypoxic (1% O-2) treatment in comparison with numerous detached dead cells in the control pcDNA3.1-transfected cells. Furthermore, a significant cell viability difference between the two kinds of transfected cells during hypoxia-reoxygenation was revealed. Therefore, the data suggest that fish HO-1 might play a significant protective role in cells in response to hypoxic stress.

Molecular cloning, characterization and expression analysis of the PKZ gene in rare minnow Gobiocypris rarus

Double-stranded RNA-activated protein kinase (PKR) plays an important rote in interferon-induced antiviral responses, and is also involved in intracellular signaling pathways, including the apoptosis, proliferation, and transcription pathways. In the present study, a PKR-like gene was cloned and characterized from rare minnow Gobiocypris rarus. The full length of the rare minnow PKR-like (GrPKZ) cDNA is 1946 bp in Length and encodes a polypeptide of 503 amino acids with an estimated molecular mass of 57,355 Da and a predicted isoelectric point of 5.83. Analysis of the deduced amino acid sequence indicated that the mature peptide contains two Zalpha domains and one S_TKc domain, and is most similar to the crucian carp (Carassius auratus) PKR-like amino acid sequence with an identity of 77%. Quantitative RT-PCR analysis showed that GrPKZ mRNA expression is at low levels in gill, heart, intestine, kidney, liver, muscle and spleen tissues in healthy animals and up-regulated by viruses and bacteria. After being infected by grass carp reovirus, GrPKZ expression was up-regulated from 24 h post-injection and lasted until the fish became moribund (P < 0.05). Following infection with Aeromonas hydrophila, GrPKZ transcripts were induced at 24 h post-injection (P < 0.05) and returned to control levels at 120 h post-injection. These data imply that GrPKZ is involved in antiviral defense and Toll-like receptor 4 signaling pathway in bacterial infection. (C) 2008 Elsevier Ltd. All rights reserved.

Microcystin-induced variations in transcription of GSTs in an omnivorous freshwater fish, goldfish

The glutathione S-transferases are important enzymes in the microcystin-induced detoxication processes. In this experiment, we cloned the full-length cDNA of alpha, pi and theta-class-like glutathione S-transferase genes from goldfish (Carassius auratus Q. Their derived amino acid sequences were clustered with other vertebrate alpha, pi and theta-class GSTs in a phylogenetic tree and the goldfish GST sequences have the highest similarity with those from common carp and zebrafish. Goldfish were i.p. injected with microcystins extract at two doses (50 and 200 mu g kg(-1) BW MC-LReq) and the relative changes of the mRNA abundance in liver, kidney and intestine were analyzed by real-time PCR. The transcription of GST alpha was suppressed in both liver and intestine, but induced in the kidney. Decreased transcription of GST theta was detected in liver, kidney and intestine in the low-dose group. The transcription of GST pi was suppressed in liver and intestine post-injection in both dose groups. These results suggested that the transcription of GST isoforms varied in different ways within an organ and among organs of goldfish exposed to MCs. (C) 2008 Elsevier B.V. All rights reserved.

Phenotypic plasticity in gut length in the planktivorous filter-feeding silver carp (Hypophthalmichthys molitrix)

Phenotypic plasticity widely exists in the external morphology of animals as well as the internal traits of organs. In the present study, we studied the gut length plasticity of planktivorous filter-feeding silver carp under different food resources in large-net cage experiments in Meiliang Bay of Lake Taihu in 2004 and 2005. There was a significant difference in stocking density between these 2 years. Under a low stocking density and abundant food resources, silver carp increased their energy intake by feeding on more zooplankton. Meanwhile, silver carp adjusted their gut length to match the digestive requirements of food when exposed to different food resources. In the main growth seasons (from April to October), silver carp significantly increased their relative gut length when feeding on more phytoplankton in 2005 (p < 0.01, 9.23 +/- 1.80 in 2004 and 10.77 +/- 2.05 in 2005, respectively). There was a nearly significant negative correlation between zooplankton proportion in the diet and the relative gut length when silver carp were stocked in a high density (p = 0.112). It appears that silver carp might have evolved plasticity to change their gut length rapidly to facilitate efficient utilization of food resources. Such resource polymorphisms in the gut may be a good indication of temporal adaptation to resource conditions. Our work provided field evidence for understanding the functional basis of resource polymorphisms and the evolution of phenotypic plasticity in planktivorous filter-feeding fish.

Triplophysa lixianensis, a new nemacheiline loach species (Pisces : Balitoridae) from the upper Yangtze River drainage in Sichuan Province, South China

Triplophysa lixianensis, a new nemacheiline loach species, is described from the Min Jiang of the upper Yangtze River drainage in Sichuan Province, South China. It can be separated from all other species of Triplophysa by having a unique combination of the following characters: posterior chamber of gas bladder greatly reduced or absent; caudal peduncle columnar with a roughly round cross- section at its beginning; anterior edge of lower jaw completely exposed or uncovered by lower lip; intestine short, forming a zigzag loop below stomach; dorsal- fin origin closer to caudal- fin base than to snout tip; pelvic fin inserted anterior to dorsal- fin origin; snout length 50.6 - 57.5 % of head length; eye diameter 12.3 15.4 % of head length; caudal peduncle length 25.1 - 27.1 % of standard length; anal fin with five branched rays; lower lip greatly furrowed with two thick lateral lobes; and body smooth or scaleless.

Analysis of heavy metals of muscle and intestine tissue in fish - in Banan section of Chongqing from Three Gorges Reservoir, China

The present study was carried out to investigate contamination of heavy metals in 19 fish species from the Banan section of Chongqing in the Three Gorges, Yangtze River. The results showed that the mean concentrations of heavy metals were higher in intestine than muscle, except zinc in upper strata. In the fish inhabiting the upper strata, there were significant differences between mean concentrations of As, Cr, Cu and Hg in muscle and intestine (P <0.05). There were also significant differences between mean concentrations of Cr and Cu in muscle and intestine in the fish inhabiting middle strata. However, significant differences between mean concentrations of As, Cd, Hg, Pb and Zn were measured in fish inhabiting bottom strata in both intestine and muscle tissues (P <0.05). For the fish inhabiting different strata, the concentrations of As, Cd, Cr, Cu, Hg and Ph in muscle and intestine of the fish from bottom strata (BS) were higher than those in both upper strata (US) and middle strata (MS); whereas a higher concentration of Zn was measured in muscle and intestine from fish inhabiting upper strata. Mean metal concentrations were found to be higher in age 11 than those in age I in Coreius heterodon (2- and 1-year odl fish respectively). The overall results indicated that fish muscle in the Banan section were slightly contaminated by heavy metals, but did not exceed Chinese food standards.

Population dynamics and maturation cycle of Camallanus cotti (Nematoda : Camallanidae) in the Chinese hooksnout carp Opsariichthys bidens (Osteichthyes : Cyprinidae) from a reservoir in China

The seasonal population dynamics and maturation cycle of the nematode Camallanus cotti in the posterior intestine of Chinese hooksnout carp Opsariichthys bidens have been studied in the Danjiangkou Reservoir of the Hubei Province in central China from September 2004 to November, 2005. The overall prevalence, mean abundance and intensity of C cotti among fish sampled (n = 700 fish) were 47%, 2.29 +/- 12.38 ( +/- S.D.) and 1-307 (average 4.89 +/- 17.74), respectively. The overall sexual ratio of female to male nematodes (excluding L3 and L4 juveniles) was 1.17:1. Statistical results showed weakly positive correlations betweerl fish length and the number of nematodes per host. The dynamics of infection of the nematode exhibited significant seasonal pattern in changes in mean abundance. A similar pattern was found for changes in nematode prevalence, although this was not statistically significant. Higher levels of infection were observed among fish sampled in summer months and the lower in the winter. Neither the prevalence nor the abundance of the parasite was significantly different between male and female hosts. The pattern of frequency distribution of the parasite in the host was found to be over-dispersed throughout the sampling period. In addition, studies on the development and maturation of the parasite in O. bidens revealed that development (maturation), recruitment of the next generation, and reproduction may be continuous year-round, although reproduction may peak during the winter. (c) 2007 Elsevier B.V. All rights reserved.

Molecular identification and expression analysis of natural resistance associated macrophage protein (Nramp) cDNA from Japanese flounder (Paralichthys olivaceus)

Natural resistance associated macrophage protein (Nramp) controls partially innate resistance to intracellular parasites. Its function is to enhance the ability of macrophages to kill pathogens. However, little is known about the structure and function of Nramp in lower vertebrates such as teleosts. We have recently isolated a cDNA encoding Nramp from Japanese flounder (Paratichthys olivaceus). The full-length cDNA of the Nramp is 3066 bp in length, including 224 bp 5' terminal UTR, 1662 bp encoding region and 1180 bp 3' terminal UTR. The 1662-nt open reading frame was found to code for a protein with 554 amino acid residues. Comparison of amino acid sequence indicated that Japanese flounder Nramp consists of 12 transmembrane (TM) domains. A consensus transport motif (CTM) containing 20 residues was observed between transmembrane domains 8 and 9. The deduced amino acid sequence of Japanese flounder had 77.30%, 82.71%, 82.67%, 79.64%, 80.72%, 90.97%, 91.16%, 60.14%, 71.48%, 61.69%, 72.37% identity with that of rainbow trout Nramp alpha and beta, channel catfish Nramp, fathead minnow Nramp, common carp Nramp, striped sea bass Nramp, red sea bream Nramp, mouse Nramp 1 and 2, human Nramp 1 and 2, respectively. RT-PCR indicated that Nramp transcripts were highly abundant in spleen, head kidney, abundant in intestine, liver and gill, and less abundant in heart. The level of Nramp mRNA in embryos gradually increases during embryogenesis from 4 h (8 cell stage) to 80 h (hatched stage) after fertilization. (c) 2005 Elsevier Ltd. All rights reserved.

Amplified fragment length polymorphism analysis to identify the genetic structure of the Gymnocypris przewalskii (Kessler, 1876) population from the Qinghai Basin, China

Amplified fragment length polymorphism (AFLP) was used to analyse the genetic structure of 45 individuals of Gymnocypris przewalskii (Kessler, 1876), an endangered and state-protected rare fish species, from three areas [the Heima (HM), Buha (BH) and Shaliu rivers (SL), all draining into Qinghai Lake]. A total of 563 polymorphic loci were detected. The HM, BH and SL populations have 435, 433 and 391 loci, respectively (Zhu and Wu, 1975), which account for 77.26%, 76.91% and 69.45% of the total number of polymorphic loci of each population, respectively. The Nei indices of genetic diversities (H) of the three populations were calculated to be 0.2869 (HM), 0.2884 (BH) and 0.2663 (SL), respectively. Their Shannon informative indices are 0.4244, 0.4251 and 0.3915, respectively. Research results show that the mean genetic distance between HM and BH is the smallest (0.0511), between BH and SL is the second shortest (0.0608), and between HM and SL is the largest (0.0713), with the mean genetic distance among the three populations being over 0.05. Data mentioned above indicate that the three populations have a certain genetic differentiation. The total genetic diversity (H-t = 0.3045) and the mean value of genetic diversity within the population (H-s = 0.2786) indicate that the variations have mainly come from within the population.

Gut contents of bighead carp (Aristichthys nobilis) and the processing and digestion of algal cells in the alimentary canal

Bighead carp is one of the most important freshwater filter-feeding fish of Chinese aquaculture. In recent decades, there have been a number of contradictory conclusions on the digestibility of algae by bighead carp based on the results from gut contents and digestive enzyme analysis or radiolabelled isotope techniques. Phytoplankton in the gut contents of bighead carp (cultured in a large net cage in Lake Donghu) were studied during March-May. In biomass, the dominant phytoplankters in the fore-gut contents were the centric diatom Cyclotella (average 54.5%, range 33.8-74.3%) and the dinoflagellate Cryptomonas (average 22.8%, range 6.8-55.8%). Phytoplankton in water samples were generally present in proportionate amounts in samples from the fore-guts of bighead carp. The size of most phytoplankton present in the intestine of bighead carp was between 8 and 20 mum in length. Bighead carp was also able to collect particles (as small as 5-6 mum) much smaller than their filtering net meshes, suggesting the importance of mucus in collecting small particles, Examination of the change in the integrity of Cyclotella on passage through the esophagus of bighead carp indicated that disruption of the algal cell walls is principally by the pharyngeal teeth, explaining the previous contradictory conclusions. (C) 2001 Elsevier Science B.V. All rights reserved.

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.

Temperature Sensitivity Dependence on Cavity Length in p-Type Doped and Undoped 1.3-mu m InAs-GaAs Quantum-Dot Lasers

We have fabricated 1.3-mu m InAs-GaAs quantum-dot (QD) lasers with and without p-type modulation doping and their characteristics have been investigated. We find that introducing p-type doping in active regions can improve the temperature stability of 1.3-mu m InAs-GaAs QD lasers, but it does not, increase the saturation modal gain of the QD lasers. The saturation modal gain obtained from the two types of lasers is identical (17.5 cm(-1)). Moreover, the characteristic temperature increases as cavity length increases for the two types of lasers, and it improves more significantly for the lasers with p-type doping due to their higher gain.

Effects of waveguide length and pump power on the efficiency of wavelength conversion in silicon nanowire waveguides

We point out the use of a wrong definition for conversion efficiency in the literature and analyze the effects of the waveguide length and pump power on conversion efficiency according to the correct definition. The existence of the locally optimal waveguide length and pump power is demonstrated theoretically and experimentally. Further analysis shows that the extremum of conversion efficiency can be achieved by global optimization of the waveguide length and pump power simultaneously, which is limited by just the linear propagation loss and the effective carrier lifetime. (C) 2009 Optical Society of America

Mode characteristics of semiconductor equilateral triangle microcavities with side length of 5-20 mu m

Semiconductor equilateral triangle microresonators (ETRs) with side length of 5, 10, and 20 mum are fabricated by the two-step inductively coupled plasma (ICP) etching technique. The mode properties of fabricated InGaAsP ETRs are investigated experimentally by photoluminescence (PL) with the pumping source of a 980-nm semiconductor laser and distinct peaks are observed in the measured PL spectra. The wavelength spacings of the distinct peaks agree very well with the theoretical longitudinal mode intervals of the fundamental transverse modes in the ETRs, which verifies that the distinct peaks are corresponding to the enhancement of resonant modes. The mode quality factors are calculated from the width of the resonant peaks of the PL spectra, which are about 100 for the ETR with side length of 20 mum.

Effect of inter-level relaxation and cavity length on double-state lasing performance of quantum dot lasers

Double-state lasing phenomena are easily observed in self-assembled quantum dot (QD) lasers. The effect of inter-level relaxation rate and cavity length on the double-state lasing performance of QD lasers is investigated on the basis of a rate equation model. Calculated results show that, for a certain cavity length, the ground state (GS) lasing threshold current increases almost linearly with the inter-level relaxation lifetime. However, as the relaxation rate becomes slower, the ratio of excited state (ES) lasing threshold current over the GS one decreases, showing an evident exponential behavior. A relatively feasible method to estimate the inter-level relaxation lifetime, which is difficult to measure directly, is provided. In addition, fast inter-level relaxation is favorable for the GS single-mode lasing, and leads to lower wetting layer (WL) carrier occupation probability and higher QD GS capture efficiency and external differential quantum efficiency. Besides, the double-state lasing effect strongly depends on the cavity length. (c) 2007 Elsevier B.V. All rights reserved.