Metallothionein induction in early larval stages of tilapia (Oreochromis mossambicus)

Amounts of whole-body metallothionein (MT) in tilapia (Oreochromis mossambicus) larvae increased to a peak (1,500 ng mg(-1) protein) 1 d after hatching (H1), decreased rapidly thereafter, and was maintained at a constant level (700 ng mg(-1)) 3 d after hatching (H3). Waterborne Cd(2+) could stimulate MT expression in newly hatched (H0) larvae in dose-dependent and time-dependent patterns. H0 larvae, which were treated with 35 microg L(-1) Cd(2+) for 24 h, showed a 1.7-fold increase in the MT amount (174.0+/-64.7) and a 6. 5-fold increase in accumulated Cd(2+) but no significant change in Ca(2+) content, compared with the H0 control (MT, 102.6+/-48.1). H3 larvae with the same treatment revealed about a 10-fold increase in accumulated Cd(2+), a 10% decrease in Ca(2+) content, but no change in MT (261.2+/-120.0), compared with the H3 control (MT, 330+/-74.0). H0 larvae could synthesize more MT to bind Cd(2+) for detoxification in 35 microg L(-1) Cd(2+), a dose that would not affect normal physiology or survival of H0 larvae. On the other hand, 35 microg L(-1) Cd(2+) caused H3 larvae to experience hypocalcemia, an abnormal physiological condition, in which H3 larvae could not synthesize sufficient MT, thus causing greater than 25% mortality. These results indicate for the first time that the inducibility of MT by waterborne Cd(2+) is development dependent, being correlated with inconsistent sensitivities to Cd(2+) during larval development.     

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.     

Cadmium,zinc, lead and copper inChironomus riparius (Meigen) larvae (Diptera,Chironomidae): uptake and effects

Cadmium, Zn, Pb and Cu uptake and effects in larvae ofChironomus riparius (Meigen) were studied in an integrated laboratory investigation, in which metal analyses in different instar stages, uptake kinetics and effects on development and growth were considered in three separate experiments.In short-term experiments with fourth instar larvae, it was demonstrated that only a minor portion of metals was adsorbed on the larval exoskeletons. No conclusive evidence on the uptake mechanism was found, but active uptake of trace metals seemed highly unlikely.In partial life cycle experiments, all four metals studied were readily accumulated in chironomid larvae. Uptake could be described satisfactorily utilizing a first-order one-compartment uptake model which incorporated growth. In all cases steady state conditions were approached and high uptake and elimination rate constants were estimated. Distinct differences between essential (Zn and Cu) and non-essential (Cd and Pb) metals were noted. Larval growth was significantly impaired upon exposure.Finally, long-term exposure experiments with low Cd concentrations (0.010 and 0.025 mg 1^{–1) resulted initially in growth impairment and high mortality in first instar stages, but surviving larvae restored growth and adults emerged even before control adults. In similar experiments with Zn (0.1 and 1.0 mg 1–1}), development of larvae was significantly retarded. High Zn concentrations in larvae were noted and almost no adult midges emerged.     

Effect of cadmium-feeding on tissue concentrations of elements in germ-free silkworm (Bombyx mori) larvae and distribution of cadmium in the alimentary canal

Silkworm (Bombyx mori) larvae were reared on an artificial diet containing cadmium (Cd) at concentrations of 5 and 80 micrograms/g wet diet from hatching to the fourth instar and then for 5 days at the fifth instar, respectively. Concentrations of Cd and other elements in the alimentary canal, Malpighian tubes, silk gland, fat body and other organs were determined simultaneously by inductively coupled argon plasma-atomic emission spectrometry. Cd was accumulated in the alimentary canal and Malpighian tubes at concentrations of 1100 and 470 micrograms/g dry wt, respectively. The distribution of Cd in the supernatants of the two highly accumulated organs were determined on an SW column by high performance liquid chromatography-atomic absorption spectrophotometry. Cd was primarily bound to inducible high molecular weight Cd-binding proteins.     

The timing of grain Cd accumulation in rice plants: the relative importance of remobilisation within the plant and root Cd uptake post-flowering

The pathways by which Cd is accumulated in rice grain are not well understood, in particular the components attributable to direct transfer from the root, and to remobilisation of Cd previously accumulated in other plant parts. In order to observe the timing of Cd accumulation in rice plants and determine the major period for accumulation of Cd which can be translocated to the grain, Cd was supplied to the roots of rice plants grown under static hydroponic conditions at a non-toxic, environmentally relevant concentration (50 nM), according to three different timing regimes: (1) Pre-flowering Cd, (2) Post-flowering Cd, or (3) Continuous Cd. The rate of accumulation of Cd in the developing grain was monitored by harvesting immature rice panicles at four time points prior to a final harvest. Nearly all grain Cd was accumulated within 16 days of anthesis and the contribution of post-flowering Cd uptake was evident from 7 days after flowering. It was estimated that 60% of the final grain Cd content was remobilised from that accumulated by the plant prior to flowering and the other 40% came from uptake during grain maturation. This study shows that Cd uptake from the root to the grain in rice is indeed possible post-flowering and it is an important source of grain Cd.     

The multiplication and spread of sacbrood virus of bees

Sacbrood virus multiplied without causing symptoms in adult bees when it was injected into them or when it was fed to young individuals. More virus accumulated in heads of infected bees than elsewhere in their bodies, and much was in their hypopharyngeal glands. The extract of each infected head contained about 10²medial lethal doses (LD 50s) of sacbrood virus for larvae when given in their food. The infective dose of sacbrood virus by injection for adults was about 10^-4of the LD 50 in food for larvae. The infective dose by mouth for adults was about 10²LD 50s for larvae, but bees older than 4–8 days could not be infected this way. Infection did not spread between adults but is probably transmitted in nature from infected adults to larvae and back to young adults that ingest remains of larvae killed by sacbrood. The youngest adult bees infected with sacbrood ate little pollen and lived only about 3 weeks in the laboratory, as did bees receiving no pollen, whereas bees fed with ample pollen lived 9 weeks.     

Transfer of microcontaminants from sediment to chironomids, and the risk for the Pond bat Myotis dasycneme (Chiroptera) preying on them

Transfer ratios of metals, PCBs, pesticides and PAHs from the sediment to chironomid larvae and adults collected in a highly contaminated area, the Biesbosch, were studied. Metal concentrations in larvae were 0.28 (Cd), 0.02 (Cr), 0.52 (Cu), 0.06 (Hg), 0.03 (Pb), 0.32 (Zn) times those found in standard sediment, on a dry weight basis. Hg and Zn were well transported to the adult stage. Dry weight ratios of contaminant residues in adults and in larvae were 0.38 (Cd), 0.23 (Cr), 0.62 (Cu), 1.03 (Hg), 0.08 (Pb), 0.94 (Zn). For PCBs and pesticides, the concentration ratios of chironomid larvae fat to sediment (dry organic matter) varied around 3.3, which is consistent with laboratory studies. Organochlorine residues in adult fat were comparable to those in larvae lipids. However, the concentrations of non-ortho PCBs were 1.7 times higher in adults. For polycyclic aromatic hydrocarbons (PAH), larval fat:sediment (organic matter) concentration ratios ranged from 0.004 to 0.1. Adult: larvae ratios for PAHs varied between 0.2 and 0.6. For naphthalene a much higher transport ratio of 2 was found. Chironomid adults are the most important potential food source of the Pond bat, which lives in low densities in the Biesbosch. The contaminant concentrations measured in the chironomids do not exceed diet levels that are thought to be safe for mammals. However, Pond bats collected in less contaminated areas contained PCB-concentrations of 9, 33 and 76 mg kg^–1 lipid weight, which are above concentrations that cause reproduction effects on Mink.     

Cd胁迫对舞毒蛾幼虫食物利用的影响及其Cd的外泌机制

为探明植食性昆虫对受重金属胁迫的寄主植物的生理生态响应机制,本研究用Cd胁迫下银中杨的叶片饲养舞毒蛾幼虫,分析舞毒蛾幼虫对食物的利用情况以及其对Cd的排毒代谢机制.结果表明: 取食Cd胁迫下银中杨的叶片后,舞毒蛾3～6龄幼虫体内的Cd浓度和Cd含量均显著高于对照,但随着幼虫龄期增长,其体内Cd浓度显著降低,而Cd含量有不同程度的提高;舞毒蛾幼虫粪便和虫蜕中的Cd浓度均显著高于对照;舞毒蛾3～5龄幼虫的食物消耗率显著高于对照,而转化率显著低于对照;3～4龄幼虫的食物利用率均与对照差异不显著,但在5龄时显著低于对照.说明在Cd胁迫下,舞毒蛾幼虫能通过有效的排毒代谢途径将体内富集的部分Cd排出体外,且高龄幼虫的排毒代谢能力强于低龄幼虫;舞毒蛾幼虫体质量的增加会对体内的Cd浓度形成一种稀释效应;舞毒蛾幼虫能通过调整食物消耗率和转化率之间的比例,来维持其正常生长发育所需的食物利用率,但超过一定限度后仍会造成食物利用率降低.     

Maternal cadmium exposure induces mt2 and smtB mRNA expression in zebrafish (Danio rerio) females and their offspring

The present study aimed to identify the effects of maternal cadmium (Cd(2+)) exposure on the mRNA expression of mt2 (metallothionein-2) and smtB (similar to metallothionein-B) in female zebrafish (Danio rerio) and their offspring (F1 larvae). Zebrafish females were exposed to 0, 8.9, 17.8, and 35.6 μM Cd(2+) for 72 h, and their ovaries and F1 larvae were collected to measure their Cd(2+) contents and their smtB and mt2 mRNA expression. Cd(2+) contents and the mRNA expression of smtB and mt2 in F1 larvae all showed positive correlations with the maternal Cd(2+) treatment dose. The mt2 was 1.9- to 3.4-fold higher than smtB in F1 larvae. Furthermore, F1 larvae had noticeably enhanced Cd(2+) tolerance after maternal Cd(2+) treatment. These results demonstrate that maternal Cd(2+) was transferred to larval fish and induced mt2 and smtB mRNA expression to protect larva against the impacts of Cd(2+). In female ovaries, mt2 expression showed a noticeable increase after exposure to a metal environment, while smtB did not show exactly the same effect. The study can only conclude that smtB might have a much different role other than just protecting against the impacts of metals.     

Sequestration and metabolism of host-plant flavonoids by the Pale Grass Blue, Pseudozizeeria maha (Lepidoptera: Lycaenidae)

Flavonoids in the eggs, larvae, pupae and adult butterflies of Pseudozizeeria maha were identified and quantified. Isovitexin (apigenin 6- C -glucoside) was the dominant flavonoid detected in pupae and imagines, whereas saponarin (isovitexin 7- O -glucoside) was the dominant flavonoid in larvae. Saponarin, biotransformed from isovitexin by the larvae, was found to be reconverted into isovitexin at the pupal stages. The total amounts of accumulated flavonoids differed between sexes, particularly in adults where the bodies of females had a much higher flavonoid concentration than that of the males. These flavonoids were also detected in the eggs. These results indicate the possibility that flavonoids are utilized in not only wing coloration but also as a chemical defense compound against natural threats.     

Presence of a single abundant storage hexamerin in both larvae and adults of the grasshopper, Schistocerca americana

We identified a single hexameric storage protein in the grasshopper, Schistocerca americana, and monitored its abundance through the last larval instar and up until reproductive competence in adults of both sexes. This storage hexamerin, termed Schistocerca americana Persistent Storage Protein (saPSP) was the most abundant soluble protein in both larvae and adults. In both sexes, saPSP abundance started out low at the onset of the last larval instar and accumulated during feeding, peaking just prior to molting. Adults of both sexes contained significant amounts of saPSP after eclosion. In adult males, saPSP content dropped continuously after eclosion and was lowest once individuals reached reproductive maturity. In contrast, adult females depleted saPSP reserves during the first days of adulthood, but subsequently accumulated significant saPSP stores. In adult females, saPSP stores peaked just prior to the completion of egg provisioning. Given the overall patterns of abundance, saPSP has functions in both larvae and adults. In addition, the observed pattern of storage hexamerin accumulation differs from patterns of accumulation in the other known grasshoppers, Locusta migratoria and Romalea microptera, suggesting that significant functional diversity has evolved in storage hexamerins among the grasshoppers.     

Difference in relative isometallothionein ratio between adult and larva of cadmium-loaded bullfrog Rana catesbeiana

Cadmium (Cd) was injected intramuscularly into adults (male) and larvae of the bullfrog, Rana catesbeiana. The chemical forms of Cd in the livers and the effect on several essential metal levels were determined by high performance liquid chromatography-atomic absorption spectrophotometry and inductively coupled plasma-atomic emission spectrophotometry, respectively. Metallothionein which binds cadmium, zinc and copper was induced in the liver and kidney of the adult bullfrog, and the protein was a mixture of two isoforms in both tissues. Although the two isoforms were also induced in the liver of the larva (tadpole), the relative ratio was the reverse of that in the adult.     

Accumulation of cadmium by the fourth instar larva of the fly Chironomus thummi

Accumulation and effects of cadmium were investigated in Chironomus thummi larvae exposed to 10, 100 and 250 mug radiolabeled Cd/1 for up to 4 days. (1) After 4 days, average cadmium accumulation was 6.6 ng Cd/mg dry weight (10 mug Cd/1 exposure) and 177 ng Cd/mg dry weight (250 mug Cd/1 exposure). (2) Dissection studies showed that by 32 hr of exposure to both cadmium concentrations, 63.5-81.4% of accumulated cadmium was confined to the posterior midgut epithelium. Light microscope autoradiography similarly showed accumulations of cadmium in posterior midgut epithelium and smaller amounts in fat body and muscle. Little cadmium was associated with Malphigian tubules, haemocoel, anterior midgut or exoskeleton. (3) After exposure to 10 or 250 mug Cd/1, 60-75% of cadmium in ultracentrifuged homogenates of whole animals or dissected guts was associated with the resulting supernatant. When supernatants were further analyzed by gel chromatography, cadmium eluted with both a high and low molecular weight peak. The relative proportions of cadmium in the two peaks varied with concentration and length of exposure. (4) Transmission electron microscopy of posterior midgut cells from animals exposed to cadmium demonstrated frequent mitochondrial lesions. Exposure to high cadmium concentrations caused some posterior midgut cells to undergo generalized structural degeneration.     

Ecotoxicity of trace metals for chironomids

The main issue of the present study was to evaluate Cd, Zn, Pb and Cu uptake in chironomid larvae and other freshwater benthic macro-invertebrates under field conditions. Secondly, laboratory experiments were performed to relate trace metal uptake in chironomid larvae with effects on growth and development, and to assess the fate of accumulated trace metals during metamorphosis or upon predation.     

Accumulation of Cd and Pb in various body parts,organs and tissues of Lymantria dispar asiatica (Lepidoptera: Erebidae)

To investigate the accumulation patterns of Cd and/or Pb in various body parts, organs and tissues of the Asian gypsy moth (Lymantria dispar) larvae under Cd and/or Pb stress, Cd and Pb treated artificial diets were used to feed the larvae in the current study. These larval body parts/organs/tissues included the heads, integuments (body walls), alimentary canals, fat bodies and hemolymphs. Our results showed that under Cd and/or Pb stress, their accumulations in larval body parts/organs/tissues were significantly higher than those in the control, with the amounts of tested metals in the fat bodies and hemolymphs and alimentary canals being significantly higher than those in the heads and integuments. Under the single Cd (0, 7.5, 10.5 mg/kg) or Pb stress (0, 55, 90, 125 mg/kg), the accumulations of these heavy metals were positively correlated with their concentrations in diets. Under the combined stress (Cd × Pb), the Cd accumulation at the lower Pb concentrations was higher than that at higher Pb concentrations for the body parts/organs/tissues, similar effects of Cd concentrations on Pb accumulations were also recorded. These results clearly showed that Cd and/or Pb were accumulated in various body parts/organs/tissues at different levels under the heavy metal stress. And accumulations of Pb/Cd were positively correlated with their concentrations in the diets under the single stress. Under the combined stresses, Cd and Pb had synergistic effects at low concentrations whereas antagonistic effects at high concentrations. The accumulations of Cd and/or Pb in the gypsy moth larvae affects normal physiological and biochemical functions, and thus affects their growth and development.     

Remobilization of cadmium in maturing shoots of near isogenic lines of durum wheat that differ in grain cadmium accumulation.

Cadmium accumulation in grain of durum wheat (Triticum turgidum L. var. durum) represents a concern to consumers. In an effort to understand the regulation of Cd accumulation in maturing grain, the remobilization of 109Cd applied to stem and flag leaves was examined in two near-isogenic lines that differ in grain Cd accumulation. Absorbed 109Cd was primarily retained in the labelling flap (50-54% and 65-80% for stem and flag leaves, respectively). Cadmium exported from the stem flap initially (3 d) accumulated in the stem in a declining gradient towards the head. Subsequent remobilization of Cd deposited in the stem was associated with Cd accumulation in the grain. Cadmium exported from the flag leaf flap was primarily directed to the grain. Little (<1%) Cd accumulated in the glumes or rachis, and transport of Cd to shoot tissues below the flag leaf node was low (<1%). On average, 9% and 17% of absorbed 109Cd accumulated in the grain 14 d after labelling the stem and flag leaf, respectively. Irrespective of labelling position, the low Cd-accumulating isoline averaged 1.5-2-fold lower Cd accumulation per grain and Cd concentration in the grain than the high Cd-accumulating isoline. Cadmium accumulation in the grain was inversely correlated with Cd retention in the stem (stem labelled) and labelling flap (flag leaf labelled) for both isolines. Cadmium translocation to the grain was not inhibited by Zn when both were applied simultaneously (50 pM 109Cd; 0.5 microM 65Zn) to the flag leaf. These results show that elevated remobilization of Cd from the leaves and stem to the maturing grain may be partially responsible for the high accumulation of Cd in durum wheat grain.     

Developmental stage-specific expression and tissue distribution of pierisin-1, a guanine-specific ADP-ribosylating toxin, in Pieris rapae

The cabbage butterfly (Pieris rapae) produces pierisin-1, an apoptosis-inducing protein against mammalian cells. In order to clarify the biological role of pierisin-1 in P. rapae, its expression during developmental stages was examined. Low levels of pierisin-1 mRNA and protein were detected in first-instar larvae. During growth until the fifth-instar larval stage, the amounts of the mRNA steadily increased to reach about 50-100 times the initial level. Then it rapidly decreased before pupation. The levels of mRNA in the pupae and the adults were as low as in the first-instar larvae. Levels of pierisin-1 protein also increased around 100 times from the first-instar to the fifth-instar larvae and then gradually decreased by over 90% during the pupal stage. Immunostaining of pierisin-1 demonstrated the protein to be mainly located in fat bodies of fifth-instar larvae and early-phase pupae. Although the staining intensity was low, fat bodies of early instars of the larvae and adults were also found to be positive. Moreover, examination of isolated fat body and other tissue samples of the insects were consistent with the above observations. Thus, the results indicate that mRNA of pierisin-1 was highly expressed in late stages of larvae, and that the protein accumulated in fat bodies where it persists during pupation.     

Cadmium is deposited in the gut content of larvae of the beetle Tenebrio molitor and involves a Cd-binding protein of the low cysteine type

Binding of cadmium (Cd) to metallothionein (MT) and non-MT proteins with low contents of cysteine has been observed in terrestrial arthropods. We recently isolated a Cd-binding protein with no cysteine that was induced in Cd-exposed larvae of the beetle Tenebrio molitor. In this study we have examined the molecular distribution of Cd within extracts of different tissues and compartments of Cd-exposed T. molitor larvae. A Cd-peak consistent with the low cysteine Cd-binding protein was induced within the gut content where it could be detected after 4-8 days of exposure. Examination of gut wall tissue revealed no increase in Cd-binding capacity, indicating that no accumulation of MTs was taking place in this tissue. Incorporation of Cd in the gut wall tissue stabilized after 8 days of Cd-exposure at a rather low level compared to the other organs. There was a statistical trend towards Cd being incorporated in the gut content in a manner that was disproportionally high compared to the amount of Cd in the gut wall tissue. The possible role of the low cysteine Cd-binding protein in reducing the uptake of Cd in the tissues is discussed.     

Maternal transfer of cadmium tolerance in larval Oreochromis mossambicus

Exposing female tilapia to Cd did not affect reproduction and egg quality, nor the survival of their offspring. The LT ₅₀ of Cd-exposed larvae (within 24 h post hatching) from Cd-treated female Oreochromis mossambicus (mean ±S.E.=10·9±0·5 days, n =4) was greater than that of Cd-exposed larvae from control females (6·9± 1·2 days, n =4). A substantial amount of metallothionein mRNA (MT mRNA) was found in both oocytes and newly hatched larvae from Cd-treated females, but MT mRNA levels were low in newly fertilized eggs. The amount of MT mRNA was very low in all these stages in the control group. Cadmium could not be detected at any stage for either control or Cd-treated group, and metallothionein protein levels were also very low in both groups. The early appearance of MT mRNA in oocytes and newly hatched larvae from Cd-treated females may result in early translation after hatching and explain their tolerance to Cd.