病毒感染草鱼胸腺的EST分析和免疫相关基因的鉴定

以感染草鱼(Ctenopharyngodon idellus)出血病病毒(GCHV)的草鱼胸腺为材料,构建了草鱼胸腺的SMARTcDNA文库.筛选文库获得到1933条有效EST序列.BLASTX分析显示,583条序列在公共数据库中能找到同源基因(E-value≤1.00E 10-3,Identities≥30%),另外1350条序列则找不到显著同源性.已知基因按具体功能可划分为6类,大部分与细胞内的各种生理过程、细胞结构以及免疫防御相关.研究结果从分子水平上表明鱼类的胸腺在机体感染病毒的免疫反应中发挥重要作用,同时也表明胸腺组织在病毒感染后可能表达很多目前还不清楚功能的新基因.     

Effects of cadmium on the reproductive axis of Japanese medaka (Oryzias latipes)

Cadmium (Cd) is a ubquitous element and a significant inorganic pollutant that has previously been found to bioaccumulate in reproductive organs of fish and disrupt important endocrine processes, especially those involved in synthesis, release and metabolism of hormones. Clearly, there is potential for reproductive effects in fish populations exposed to Cd, however, few studies have investigated the non-lethal consequences of Cd in fish. To this extent, adult male and female Japanese medaka were exposed to 0-10 ppb Cd for 7 weeks. Reproductive endpoints were monitored during weeks 6 and 7 of exposure and compared to physiological responses along the hypothalamus-pituitary-gonadal (HPG) axis, including plasma vitellogenin (VTG), hepatic estrogen receptor (ER), plasma steroids, gonadal-somatic indices (GSI), and gonadal steroid release. There were no observed effects on VTG and ER by long-term Cd exposure. However, gonadal steroid release was significantly decreased in males and females at all exposure concentrations and female plasma estradiol levels were significantly altered at concentrations higher than 5 ppb Cd. Overall, responses along the HPG axis were more sensitive to Cd exposure than the reproductive and developmental endpoints, which were not affected in this study, indicating that higher level impairment in fish might be relatively protected.     

Toxicological stress response and cadmium distribution in hybrid tilapia (Oreochromis sp.) upon cadmium exposure

Adult tilapia were exposed to 0 (control) and 4.45 microM Cd for 0 h, 5 h, 5 days, and 15 days, and the physiological responses of fish were described. The physiological responses were first expressed in gill tissue, in which mucus cells secretion increased, Cd accumulated, cortisol secretion was significantly higher, but serum ACH(50) activity (alternative complement hemolytic assay) was significantly lower than in controlled fish. After 5 days of Cd exposure, the ACH(50) activity showed a greater decrease, but lysozyme and cortisol contents showed significant increases over the control. Cd levels significantly increased in intestines, liver, and kidneys, and a significant induction of metallothionein (MT) protein in hepatic tissue was noted. Finally, the Cd accumulation rate still showed significant increases in these organs. However, the MT content was similar at 5 days and at 15 days after Cd exposure, and the cortisol contents had recovered to the pre-exposure level. In addition, Cd accumulation in muscle was higher after Cd exposure than in controls (t-test, p<0.05). Our results demonstrate (1) that tilapia readily regulate their physiological parameters in order to acclimate to a sublethal Cd environment; (2) these changes of physiological parameters may be related with a succession of cortisol levels following Cd exposure; (3) increasing rate of hepatic MT contents and Cd accumulation rate didn't appear identical after 5-15 days of Cd exposure. This was confirmed that hepatic MT was not a good indicator for Cd levels in tilapia.     

Effects of Bothriocephalus acheilognathi on the polarization response of pronephric leucocytes of carp, Cyprinus carpio

An in vitro assay was used to examine the effect of Bothriocephalus acheilognathi Yamaguti, 1934 (Cestoda: Pseudophyllidea) on the polarization response of pronephric leucocytes of carp, Cyprinus carpio. Leucocytes, isolated from naive, naturally-infected fish and carp injected intraperitoneally with cestode extracts, were exposed to parasite extracts (protein concentrations 0-10.0 microg ml-1), for up to 24 h in the presence or absence of carp serum. In general, polarization responses of the pronephric leucocytes, primarily neutrophils and eosinophils, increased with incubation time although there was no significant difference in the response induced by the different protein concentrations. Differences in the polarization response were, however, observed in naive, naturally infected and injected fish and the cells responded differently in the presence and absence of carp serum. In the absence of carp serum the polarization response of pronephric leucocytes in vitro was significantly reduced with cells obtained from injected and naturally infected fish compared with those obtained from naive carp. This suppression of leucocyte migration was however reduced by the addition of carp serum to the in vitro system. The role of this interaction between the possible suppression of polarization induced by the parasite and stimulation by serum is discussed.     

Effects of Montmorillonite on Alleviating Dietary Cd-Induced Oxidative Damage in Carp (Carassius auratus)

The present study was designed to investigate the effects of montmorillonite (MMT) on dietary Cd-induced oxidative damage in liver and kidney of carp (Carassius auratus). One hundred eighty carp were randomly divided into four groups and fed with a basal diet, a basal diet supplemented with 0.5% MMT, Cd-comtaminated basal diet (120 mg Cd/kg dry weight) and Cd-contaminated basal diet supplemented with 0.5% MMT, respectively. After 60 days, fish were sacrificed to measure malondialdehyde (MDA) content and antioxidative indices in liver and kidney. The results showed that the exposure of carp to dietary Cd caused decreases in glutathione peroxidase activity, catalase activity, superoxide dismutase activity, glutathione content and total antioxidant capacity level, while MMT supplemented in diet compensated Cd-induced decreases in above antioxidant indices to some extent in liver and kidney. As compared with the control group, increases in MDA content were observed in both measured tissues of carp exposed to dietary Cd, while MDA content decreased in carp exposed to Cd-contaminated basal diet supplemented with MMT in comparison with the Cd-contaminated group. It was suggested that MMT, when co-administered with Cd in diet, could alleviate dietary Cd-induced oxidative damage in liver and kidney of carp.     

Effects of Cadmium Exposure on Growth and Metabolic Profile of Bermudagrass [Cynodon dactylon (L.) Pers.]

Metabolic responses to cadmium (Cd) may be associated with variations in Cd tolerance in plants. The objectives of this study were to examine changes in metabolic profiles in bermudagrass in response to Cd stress and to identify predominant metabolites associated with differential Cd tolerance using gas chromatography-mass spectrometry. Two genotypes of bermudagrass with contrasting Cd tolerance were exposed to 0 and 1.5 mM CdSO₄ for 14 days in hydroponics. Physiological responses to Cd were evaluated by determining turf quality, growth rate, chlorophyll content and normalized relative transpiration. All these parameters exhibited higher tolerance in WB242 than in WB144. Cd treated WB144 transported more Cd to the shoot than in WB242. The metabolite analysis of leaf polar extracts revealed 39 Cd responsive metabolites in both genotypes, mainly consisting of amino acids, organic acids, sugars, fatty acids and others. A difference in the metabolic profiles was observed between the two bermudagrass genotypes exposed to Cd stress. Seven amino acids (norvaline, glycine, proline, serine, threonine, glutamic acid and gulonic acid), four organic acids (glyceric acid, oxoglutaric acid, citric acid and malic acid,) and three sugars (xylulose, galactose and talose) accumulated more in WB242 than WB144. However, compared to the control, WB144 accumulated higher quantities of sugars than WB242 in the Cd regime. The differential accumulation of these metabolites could be associated with the differential Cd tolerance in bermudagrass.     

Potentiation of autoimmune response in rats infected with Toxoplasma gondii. Effect of the infection route

The aim of this report is to describe the influence of the way of infection with 3 X 10(3) trophozoites of Toxoplasma gondii on the auto- and heteroimmune responses of rats immunized with chemically modified rat male accessory glands (MRAG) and human serum albumin (HSA) 6 and 36 days after infection. The delayed hypersensitivity (DTH) response to MRAG was potentiated in rats infected in thymus proximity (p less than 0.02) when compared with rats only immunized. The parasites introduced intraperitoneally did not modify the cellular autoimmune response. The titers of hemagglutinating antibodies to MRAG were increased in animals infected by both ways (p less than 0.05). The response to HSA did not show significant difference between the infected and uninfected rats. Thymic involution and proliferation of lymphocytes and plasma cells were observed at 6 and 52 days post-infection only in rats infected in thymus proximity (p less than 0.05). A correlation between the potentiation of cellular autoimmune response and the injection of parasites in thymus proximity was observed.     

Analysis of cadmium and lead in mice organs

Analysis and distribution of Pb and Cd in different mice organs, including the liver, kidney, spleen, heart, and blood, were evaluated before and after treatment with different aqueous concentrations of Nigella sativa (1.25–10.0 mg/L). Atomic absorption spectrometry was used for analysis of Pb and Cd in these organs. Results indicated that the Pb in the unexposed group of mice without treatment with N. sativa (black cumin) was in the following order: liver>heart>spleen>kidney, and the distribution of Pb in various organs of the unexposed group was not affected significantly by N. sativa. Moreover, results of mice exposed for Pb show that the Pb concentrations in different organs were reduced significantly (p<0.05) by 72.9%, 63.4%, 72.3%, 66.7%, and 39.5% at a dose of 10 mg/L of N. sativa for the liver, kidney, heart, spleen, and blood, respectively. Furthermore, the distribution of Cd in the unexposed Cd group of mice without treatment with N. sativa was in the following order: kidney>heart>spleen>liver. Nigella sativa at 10 mg/L reduced Cd levels in mice exposed to Cd by 75.5%, 83.3%, 47.0%, 95.3%, and 100% in the liver, kidney, heart, spleen, and blood, respectively, whereas blood Cd concentrations were lowered to below the detection limit of 0.05 μg/L. A 28-d exposure of mice to a Cd−Pb mixture at a concentration of 1 ppm in drinking water induced a highly significant inhibition (p<0.0001) of antibody response to human serum (80.5%). The suppressed immune responses in mice pretreated with the Cd−Pb mixture were reversed by 43.1% and 38.9% in the presence of 1.25 and 2.5 mg/mL of N. sativa, respectively, whereas higher concentrations (5–10 mg/mL) of N. sativa increased the immunosuppression significantly. Nigella sativa at 1.25–10 mg/mL did not induce any significant modulation of the antibody response in unexposed mice.     

Cadmium toxicity and snail-digenean interactions in a population of Lymnaea spp

The toxicity of cadmium to a population of Lymnaea peregra and L. stagnalis naturally infected with a range of digeneans and collected from a number of sites in the lower Thames Valley, UK was investigated. Lymnaeid snails were exposed to 100 microg l-1 cadmium and the effects on host survival and emergence of cercariae recorded. Overwintered L. peregra, but not L. stagnalis, showed significantly reduced survival compared to seasonally infected snails, i.e. snails which have acquired an infection during the spring or summer. A significant increase in survival with increasing snail size was demonstrated for L. stagnalis and for seasonally infected L. peregra only. Only L. stagnalis infected with Diplostomum spathaceum and L. peregra infected individually with D. spathaceum, Sanguinicola inermis, Echinoparyphium recurvatum and Notocotylus attenuatus demonstrated a significantly reduced survival compared to laboratory-bred controls. The exposure of L. stagnalis to cadmium resulted in a significant reduction in the emergence of D. spathaceum over a 5-day period but cadmium-exposed L. peregra showed no difference in the emergence of E. recurvatum cercariae over a 3-day exposure period. The mechanisms and importance of metal toxicity to snail-digenean interactions are discussed.     

Accumulation of cadmium and its effects on growth, development and hemolymph biochemical compositions in Boettcherisca peregrina larvae （Diptera： Sarcophagidae）

Newly emerged larvae of the fleshfly, Boettcherisca peregrina were exposed to two different CdCl2 concentrations of 100μg/g and 400 mg/g diet fresh weight （DFW）. They were administered in the diets until the end of larval stage. Cd-exposed larvae accumulated significant amounts of Cd and this accumulation increased with the exposure dose and time. The body weights were lightened and lengths of larvae were shortened considerably after Cd exposure, especially at the higher Cd concentration. The total larval duration was also extremely affected due to Cd exposure. The average duration was prolonged significantly by 14 h at the lower Cd concentration, while it was increased by 33.7 h over controls at the higher Cd concentration. A significant decrease in contents of either soluble proteins, total lipids or caloric values in the hemolymph occurred due to Cd exposure throughout the entire tested period but after 120 h of Cd exposure. In contrast, when exposed to Cd with its higher concentration, total sugar contents in the hemolymph were increased strikingly over the whole tested time, except after 96 h of Cd exposure, while they were not apparently altered except after 24 h of Cd exposure at the lower concentration. Thus, it is suggested that Cd exposure shows significant adverse impact on the growth and development, as well as metabolism, in larvae of this fleshfly, depending on its exposure time and dose.     

氨氮胁迫下不同食性鱼类仔鱼抗氧化和非特异性免疫的差异响应

为探究氨氮胁迫对不同食性鱼类的影响,研究选取不同食性的4种鱼类(鲢Hypophthalmichthys molitrix、草鱼Ctenopharyngodon idella、团头鲂Megalobrama amblycephala和黄颡鱼Pelteobagrus fulvidraco)仔鱼为研究对象,探讨不同浓度氨氮(0、1、2和3 mg/L)短期胁迫(96h)对其生长、抗氧化和非特异性免疫响应的影响及其分子机制。结果显示:(1)氨氮胁迫导致4种仔鱼的体长生长速度呈现剂量依赖性减缓;(2)不同浓度氨氮导致4种仔鱼体内T-AOC、CAT和GPx含量显著降低(P<0.05), 2和3 mg/L氨氮暴露显著降低了草鱼、团头鲂和黄颡鱼仔鱼体内SOD活力(P<0.05),仅检测到黄颡鱼gpx及鲢sod转录水平出现显著性下调(P<0.05);(3)在不同浓度氨氮胁迫下, 4种仔鱼相关免疫基因转录水平均呈现一定的上调,仅鲢il1β转录水平显著下降(P<0.05),相对地草鱼仔鱼LYZ含量显著性下降,黄颡鱼仔鱼LYZ含量在2 mg/L氨氮组显著性上升(P<0.05)。双因素...     

Environmental Cadmium Exposure Impacts Physiological Responses in Manila clams

The physiological responses of marine bivalves to chronic cadmium (Cd) exposure at sub-lethal concentrations have been well documented. As of now, few studies have examined the effect of Cd exposure and subsequent recovery period at environmentally realistic concentrations. In this study, environmentally, Cd exposures were performed to assess the physiological responses of the Manila clam Ruditapes philippinarum. The clams were exposed to waterborne Cd at two environmentally realistic concentrations (4 and 40 μg L^?1) for 35 days and then allowed to recover for another 35 days. The accumulation and elimination of Cd in R. philippinarum were tissue-specific and dose- and time-dependent. Cd accumulation increased sharply in the digestive gland, and Cd elimination was rapid in the gill. Major physiological responses, including clearance rate, absorption efficiency, respiration rate, excretion rate, oxygen to nitrogen ratio, and scope for growth, were significantly affected by Cd exposure. Yet, the clams exposed to 4-μg L^?1 Cd were able to quickly recover their normal physiological processes and clearly exhibited catch-up growth once they were transferred to clean seawater. Hence, R. philippinarum can exhibit good physiological plasticity when confronted with moderately environmental Cd exposure. All physiological responses measured exhibited a highly significant and generally predictable correlation with tissue Cd concentration, which in turn, reflected environmentally realistic exposure conditions. Our results further confirm that the measurement of physiological responses is a sensitive method for assessing stress at environmentally realistic metal concentrations.     

Net cadmium flux and accumulation reveal tissue-specific oxidative stress and detoxification in Populus × canescens

To characterize the dynamics of Cd²⁺ flux in the rhizosphere and to study cadmium (Cd) plant‐internal partitioning in roots, wood, bark and leaves in relation to energy metabolism, reactive oxygen species (ROS) formation and antioxidants, Populus × canescens plantlets were exposed to either 0 or 50 µM CdSO₄ for up to 20 days in the nutrient solution. A strong net Cd²⁺ influx in root apex was observed after Cd exposure for 24 h, even if net Cd²⁺ influx decreased gradually in roots. A large amount of Cd was accumulated in roots. Cd ions were uploaded via the xylem to leaves and further transported to the phloem where significant accumulation was detected. Cd accumulation led to decreased photosynthetic carbon assimilation but not to the depletion in soluble carbohydrates. Increased levels of ROS were present in all tissues, except the bark of Cd‐exposed poplars. To combat Cd‐induced superoxide and hydrogen peroxide, P.×canescens appeared to rely mainly on the formation of soluble phenolics as these compounds showed the highest accumulation in the bark and the lowest in wood. Other potential radical scavengers such as proline, sugar alcohols and antioxidant enzymes showed tissue‐ and exposure time‐specific responses to Cd. These results indicate a complex pattern of internal Cd allocation in P.×canescens resulting in higher ROS stress in wood than in bark and intermediate responses in roots and leaves, probably because of differential capacities of these tissues for the production of protective phenolic compounds.     

The thymus is a common target organ in infectious diseases

Infectious disease immunology has largely focused on the effector immune response, changes in the blood and peripheral lymphoid organs of infected individuals, and vaccine development. Studies of the thymus in infected individuals have been neglected, although this is progressively changing. The thymus is a primary lymphoid organ, able to generate mature T cells that eventually colonize secondary lymphoid organs, and is therefore essential for peripheral T cell renewal. Recent data show that normal thymocyte development and export can be altered as a result of an infectious disease. One common feature is the severe atrophy of the infected organ, mainly due to the apoptosis-related depletion of immature CD4+CD8+ thymocytes. Additionally, thymocyte proliferation is frequently diminished. The microenvironmental compartment of the thymus is also affected, particularly in acute infectious diseases, with a densification of the epithelial network and an increase in the deposition of extracellular matrix. In the murine model of Chagas disease, intrathymic chemokine production is also enhanced, and thymocytes from Trypanosoma cruzi-infected mice exhibit greater numbers of cell migration-related receptors for chemokines and extracellular matrix, as well as increased migratory responses to the corresponding ligands. This profile is correlated with the appearance of potentially autoreactive thymus-derived immature CD4+CD8+ T cells in peripheral organs of infected animals. A variety of infectious agents--including viruses, protozoa, and fungi--invade the thymus, raising the hypothesis of the generation of central immunological tolerance for at least some of the infectious agent-derived antigens. It seems clear that the thymus is targeted in a variety of infections, and that such targeting may have consequences on the behavior of peripheral T lymphocytes. In this context, thymus-centered immunotherapeutic approaches potentially represent a new tool for the treatment of severe infectious diseases.     

Analysis of cadmium and lead

Chronic exposure (3.5 mo) of mice to cadmium (Cd), lead (Pb), or a cadmium-lead mixture at a concentration of 1 ppm in drinking water induced a highly significant inhibition of antibody response to human serum. The highest immunosuppression (84.4%) was induced by the Cd-Pb mixture, whereas Cd caused the lowest immunosuppression (53.6%). The body burden of Cd and Pb in various organs was investigated in the four groups of mice by atomic absorption spectrometry. The highest level of Cd was found in the kidney of the Cd-treated group, and the highest level of Pb was found in the liver of the Pb- and Cd-Pb-treated groups. It is concluded that when mice are exposed concurrently to Cd and Pb, they develop synergistic immunsuppression. Analysis of Cd levels using atomic absorption spectrometry revealed that it was distributed in the following order: kidney>liver>spleen>heart, whereas Pb was distributed in the following order: liver>kidney>spleen>heart.     

Analysis of cadmium and lead: their immunosuppressive effects and distribution in various organs of mice

Chronic exposure (3.5 mo) of mice to cadmium (Cd), lead (Pb), or a cadmium-lead mixture at a concentration of 1 ppm in drinking water induced a highly significant inhibition of antibody response to human serum. The highest immunosuppression (84.4%) was induced by the Cd-Pb mixture, whereas Cd caused the lowest immunosuppression (53.6%). The body burden of Cd and Pb in various organs was investigated in the four groups of mice by atomic absorption spectrometry. The highest level of Cd was found in the kidney of the Cd-treated group, and the highest level of Pb was found in the liver of the Pb- and Cd-Pb-treated groups. It is concluded that when mice are exposed concurrently to Cd and Pb, they develop synergistic immunosuppression. Analysis of Cd levels using atomic absorption spectrometry revealed that it was distributed in the following order: kidney > liver > spleen > heart, whereas Pb was distributed in the following order: liver > kidney > spleen > heart.     

Formation of metallothionein in fish

1. Carp were bred in Cd solution, and each organ was homogenized and centrifuged at 105,000 g. The Cd content of the supernatant was much more than that of the precipitate. 2. In examining the metal form in the supernatant, the induction of metallothionein (MT) was found in carp exposed to Cd and Zn solutions and the presence of metallothionein was found in fish captured in Nagara river and breeding ponds. 3. Cd content in the metallothionein fraction of hepato-pancreas increased at the early stages of exposure, but Cd in the metallothionein fraction of the other organs increased as time proceeded. 4. In the tap water group, metallothionein in hepato-pancreas and kidney contained Cu, and increased with the induction of metallothionein in hepato-pancreas due to Zn-exposure.     

Multi-biomarker responses in the freshwater mussel Dreissena polymorpha exposed to polychlorobiphenyls and metals

Contaminant related changes in behavioral, phase I and II metabolizing enzymes and pro-oxidant/antioxidant processes in the freshwater mussels Dreissena polymorpha exposed to metals and PCBs were assessed. Behavioral and biochemical responses including filtering rates, key phase I, II and antioxidant enzymes and levels of metallothioneins, glutathione, lipid peroxidation and DNA strand breaks were determined in digestive glands of mussels after being exposed to sublethal levels of mercury chloride, methyl mercury, cadmium and Aroclor 1260 during 5 days. In 7 out of 12 responses analyzed, mussels showed significant differences across treatments. Unusual properties of measured ethoxyresorufin-O-deethylase (EROD) activities indicated that mussels lack an inducible CYP1A enzymatic activity. Despite of using similar exposure levels, inorganic and organic mercury showed different biomarker patterns of response with methyl mercury being more bio-available and unable to induce metallothionein proteins. Mussels exposed to Cd presented higher levels of metallothioneins and an enhanced metabolism of glutathione, whereas those exposed to Aroclor showed their antioxidant glutathione peroxidase related enzyme activities inhibited. Although there was evidence for increased lipid peroxidation under exposure to inorganic and organic mercury, only mussels exposed to Aroclor had significant greater levels than those in controls.     

Cadmium exposure of rainbow trout, Salmo gairdneri Richardson: effects on immune functions

Differential leucocyte counts, phagocytosis, humoral antibody response and the in vitro blasto-genetic response to mitogens (lipopolysaccharide and Concanavalin A) and to an antigen ( Vibrio anguillarum ) were studied in rainbow trout exposed to 0,0.7 or 3.6 μg Cd 1⁻¹ for 12 weeks.
Although the fish did not exhibit any clinical or histological changes, cadmium exposure was found to affect two of the immune parameters measured. The cellular response of fish immunized with V. anguillarum to the homologous antigen was significantly lower for splenocytes obtained from fish exposed to cadmium for 9 weeks (3.6 μg Cd 1⁻¹ group) than for splenocytes obtained from non-exposed fish. Conversely, the humoral antibody response to V. anguillarum O-antigen was higher in the 3.6 μg Cd 1⁻¹ group than in the non-exposed group. Protective immunity of fish vaccinated against V. anguillarum was equally as good in the cadmium-exposed group as in the non-exposed group. No cadmium-induced changes in differential leucocyte counts or in the proportions of phagocytic cells were observed.