Effects of zinc and female aging on nymphal life history in a grasshopper from polluted sites

Insect reproduction is influenced by various factors, including food quality and quantity, temperature, population density and female age. Contamination, including heavy metals, may disturb reproductive processes. The aim of this work was to assess interactions between effects of aging in female Chorthippus brunneus and environmental pollution on their reproduction measured in number of laid eggs. We also compared basic developmental parameters (number of hatchlings, body mass, embryonic developmental rate) in grasshopper nymphs additionally exposed to zinc during diapause. Aging grasshoppers from heavily polluted areas (Olkusz and Szopienice) lay significantly fewer eggs than insects from the reference site (Pilica). Zinc application caused the decrease in hatching success and duration of embryogenesis in insects from each site. This suggests a cumulative effect of female age, pollutants and additional stressing factors. The intensity of this process differed between populations. In insects from the reference site, it was shown in a moderate degree. In insects from Szopienice, an additional stressor exerted a weaker effect than in insects from Pilica. In grasshoppers from Olkusz, we found the strongest decrease of hatching percentage and increase in duration of embryogenesis after zinc intoxication. This may indicate that the population from Olkusz exists at the limit of its energetic abilities.     

Oxidative stress in newly-hatched Chorthippus brunneus--the effects of zinc treatment during diapause, depending on the female's age and its origins

The responses of glutathione, glutathione S-transferases (GSTs), and catalase (CAT) were determined in 1-day-old larvae of Chorthippus brunneus Thunberg, 1815, a grasshopper exposed to zinc during diapause, from unpolluted (Pilica) or polluted (Olkusz, Szopienice) sites. The aim of the work was to search for differences among populations of the insects as a result of various multistress pressures in their habitats. The question of zinc toxicity in the context of energy allocation was also considered. Zinc caused a decrease in glutathione concentration in the body of zinc-treated larvae. Significant differences between control and zinc-treated groups were confirmed for young females' progeny from Pilica and Olkusz as well as old females' progeny from Olkusz. GSTs activity was generally not influenced by zinc. It is possible that GSTs were not the most important target of zinc action. On the contrary, the influence of zinc on CAT activity was found. The increase in CAT activity after zinc treatment was similar for all studied populations. An increase in CAT activity after zinc exposure seems to be the most universal reaction. CAT activity in zinc-treated grasshoppers may explain the mechanism of zinc toxicity based on reactive oxygen forms generation.     

Hsp70 level in progeny of aging grasshoppers from variously polluted habitats and additionally exposed to zinc during diapause

The hsp70 level in the bodies of 1st instars of grasshoppers Chorthippus brunneus from unpolluted (Pilica) and polluted (Olkusz, Szopienice) sites and additionally exposed to various doses of zinc during diapause and embryonic development prior to hatching were measured by Western blotting. The main aim of our work was to assess the relationship between the age of female grasshoppers originating from variously polluted habitat and the hsp70 level in their progeny. Possible reasons for population variation in hsp70 levels were discussed. The hsp70 level in the offspring's body depended on the place of origin. The strongest expression of hsp70 was found in the bodies of larvae hatching from the eggs laid by young females from Pilica (reference site). In contrast, a low initial level of hsp70 in larvae from polluted sites, especially in young females’ progeny, was observed. The application of zinc during diapause influenced the hsp70 level in grasshopper larvae; however, the direction of the changes depended on the insects’ place of origin. In larvae from the reference site, and also (but to a lesser degree) from Olkusz, the increase in the hsp70 level after zinc treatment was most pronounced. Whereas in grasshoppers from Szopienice, zinc (in 100 μg g⁻¹ dry weight of sand) did not change the hsp70 level, or (in 500 μg g⁻¹ dry weight of sand) caused a reduction in hsp70. The differences may result from maternal effects; however, possible adaptation also cannot be excluded. To confirm this statement further studies are needed.     

Does the grasshopper Chorthippus brunneus adapt to metal polluted habitats? A study of glutathione‐dependent enzymes in grasshopper nymphs

Glutathione contents and activity of glutathione-dependent enzymes in the body of insects inhabiting polluted areas depend on toxin kind, concentration and exposure time. Enzymatic response may be modified by gender, age, developmental stage and state of nutrition. Also, chronic exposure to metals in the environment may cause the selection of individuals resistant to some environmental toxins. To assess the degree of adaptation of Chorthippus brunneus to metal-polluted habitats, we measured glutathione contents and the activity of selected glutathione-dependent enzymes in the offspring of aging mothers which differed in time and intensity of exposure to metals in their habitats. We tested whether differences represent temporal shifts in tolerance range or were genetically preserved and inherited by future generations. We investigated insects from three populations. Two live in heavily metal-burdened areas, exposed to metals for 170 (Szopienice) or 50 years (Olkusz) and the third inhabits an unpolluted reference site (Pilica). The most important findings were age-by-site interactions for all biochemical analyses. Nymphs from Szopienice had lower glutathione contents and lower glutathione-dependent enzyme activity in comparison with nymphs from the reference site. This was especially distinct in nymphs hatched from eggs laid by young females. The offspring of aging females from Olkusz, in terms of glutathione contents and glutathione reductase activities, revealed similar patterns to those from the reference site. For the remaining parameters, enzyme activity patterns in nymphs from Olkusz were similar to those of nymphs from Szopienice.     

重金属污染影响植食性昆虫的研究进展

重金属污染是世界各国面临的最为棘手的问题之一,对生态系统和食品安全构成了严重威胁。作为生态系统中食物链和食物网的重要环节,植食性昆虫是环境中重金属迁移、积累的重要媒介,其因重金属污染而受到的影响引起了大家的关注。本文综述了从2007至2018年重金属污染对植食性昆虫影响的研究进展。昆虫受重金属胁迫的研究途径有人工饲料添加、野外田间暴露、“土壤-植物-昆虫”食物链传递以及体外注射等。积累在植食性昆虫体内的过量重金属可导致其存活率、繁殖力和种群增长率降低,生长发育迟缓。重金属污染对植食性昆虫的生理生化毒性包括细胞超微结构破坏和DNA损伤,体内能量物质含量降低,酶活性、基因表达改变等。植食性昆虫会通过重金属硫蛋白、解毒酶活性的诱导等机制抵御重金属的毒害,从而对低浓度、长期重金属暴露产生生态适应性,甚至提高对其他逆境(如农药等)的耐受性。     

Joint effects of dimethoate and heavy metals on metabolic responses in a grasshopper (Chorthippus brunneus) from a heavy metals pollution gradient

We studied how an exposure to an additional stressing factor-dimethoate, might affect detoxifying ability of grasshoppers collected at 5 meadow sites located along a heavy metal pollution gradient. Activities of esterases and enzymes linked with glutathione (GSH) metabolism were assayed 24 h after topical treatment with 0.32 microg dimethoate per insect. Inhibition of acetylcholinesterase (AChE) reaches nearly 50% of the value stated in untreated insects, without significant site-dependent differences. The pesticide also caused a significant decrease in activities of glutathione peroxidase (GPx) followed by a decrease in GSH levels in grasshoppers from all assayed groups, demonstrating high sensitivity of glutathione-dependent metabolism to the additional stressing factor. In the case of glutathione reductase (GR) and carboxylesterases (CarE) the fall of activity was shown especially in insects from less polluted meadows and the reference site. Glutathione reductase (GR) activity in individuals treated with dimethoate did not decrease only in insects from the most contaminated site I. This might suggest the trade-off mechanisms adapting grasshoppers to life in seriously polluted environments.     

Apoptosis and biochemical biomarkers of stress in spiders from industrially polluted areas exposed to high temperature and dimethoate

The aim of this study was to evaluate the relations between apoptosis and the activity of antioxidant enzymes (superoxide dismutase; catalase) and quantitative changes in stress protein positive cells (Hsp70; metallothionein) in midgut glands of funnel web spiders Agelena labyrinthica (Agelenidae) and wolf spiders Pardosa lugubris (Lycosidae) exposed to high temperature and pesticide under laboratory conditions. The spiders were collected from two meadow ecosystems differently polluted with metals (Olkusz and Pilica, southern Poland). Under stress conditions, P. lugubris had fewer apoptotic cells in the midgut glands than A. labyrinthica. In P. lugubris from both sites, the observed increase in the percentage of metallothionein and Hsp70-positive cells, simultaneous with intensification of superoxide dismutase and catalase activity, suggests an anti-apoptotic function of those proteins in representatives of wandering spiders. In the midgut glands of A. labyrinthica, heat shock and dimethoate increased the number of Annexin V-positive cells as well as the amounts of mitochondria with low transmembrane potential (DeltaPsi(m)) versus the control. The changes in the percentage of MT- and Hsp70-positive cells in funnel web spiders were less than in wolf spiders. The absence of change in SOD and CAT activity in A. labyrinthica shows that the participation of those enzymes in antioxidant reactions is minimal in this species.     

Antioxidative responses in females and males of the spider Xerolycosa nemoralis (Lycosidae) exposed to natural and anthropogenic stressors

The aim of this study was to assess the intensity of enzymatic antioxidative parameters [i.e., superoxide dismutase (SOD), catalase (CAT), and the glutathione peroxidases each selene dependent, GPOX or selene independent, including GSTPx, glutathione S-transferase, and GST] and non-enzymatic antioxidative parameters [i.e., glutathione total (GSH-t), the heat shock proteins of Hsp70, and metallothioneins (Mt)] in the midgut glands of female and male wolf spiders Xerolycosa nemoralis (Lycosidae) exposed to natural stressors (i.e., heat shock and starvation) and anthropogenic stressors (i.e., the organophosphorous pesticide dimethoate) under laboratory conditions. The spiders were collected from two differentially polluted sites both localized in southern Poland: Olkusz, which is heavily polluted with metals, and Pilica, the reference site. In response to the stressing factors, increases in Hsp70 levels, in the concentrations of total glutathione and in the activity levels of glutathione-dependent enzymes (GPOX, GSTPx, and GST) were found in the midgut glands of males. In the females, high levels of activity of CAT and SOD were revealed, as well as an increased percentage of Mt-positive cells. Preexposed females, in comparison to the individuals from the reference site, responded with increased SOD activity, irrespective of the stressing factor. In contrast, the changes in the antioxidative parameters in the midgut glands of male X. nemoralis seem to reflect a short-term reaction to the applied stressors and do not confirm the effects of long-term selection in a polluted environment.     

重金属诱导拟南芥原生质体DNA 损伤的单细胞凝胶电泳检测

以拟南芥原生质体为实验体系, 研究不同浓度的3种重金属离子对拟南芥原生质体的毒性和DNA损伤的差异。结果表明, 用1-5 mmol.L-1的Zn2+、Cd2+ 和Cu2+分别处理的拟南芥原生质体, 2 小时内活力逐渐下降, 并表现出明显的浓度依赖性;与相同浓度的Cd2+ 和Cu2+ 相比, Zn2+对拟南芥原生质体活力的影响程度较小, 表现出较低的毒性。单细胞凝胶电泳检测发现,用0.1-0.8 mmol .L-1的Zn2+、Cd2+ 和Cu2+ 分别处理拟南芥原生质体30 分钟, 以OTM值表示的原生质体DNA损伤量随重金属离子浓度的增加而递增; 相同浓度(0.5 mmol.L-1)的3种重金属离子相比, Zn2+对原生质体的遗传毒性明显低于Cu2+ 和Cd2+。综合原生质体活力和DNA损伤的单细胞凝胶电泳检测结果, 发现Zn2+对拟南芥原生质体的遗传毒性较低, 而Cd2+ 和Cu2+的遗传毒性较高。本研究建立的拟南芥原生质体实验体系, 结合运用单细胞凝胶电泳技术, 能够快速、灵敏地检测重金属对植物细胞的遗传毒性。     

A New Laboratory Technique for Studying the Effects of Heavy Metals on Bryophyte Growth

A novel, non-destructive method for measuring moss gametophytegrowth rates is presented. Standard 2-cm moss gametophores aresupported vertically on Velcro strips held within a clear perspexbox, with their bases resting in liquid. Shoots sampled fromthe field were either continuously exposed to test solutionsvia their cut bases, or by preliminary pulse-immersion. Growthwas assessed by linear extension, or final dry mass, and showeddifferent patterns and sensitivities to heavy metals using thetwo exposure methods. A biologically appropriate assessmentof toxicity was established, using tissue concentrations comparableto material from polluted field sites, by considering the cellularlocation of heavy metals in shoots. A linear relationship wasestablished between apical growth and intracellular Zn concentrationrecovered in the newly produced gametophyte tissue, irrespectiveof the method of exposure or Zn concentration in the remainingor original shoot portion. The limitations and ambiguity associatedwith many heavy-metal monitoring and toxicity studies are addressed,and the advantages and further applications offered by thisgrowth system are discussed. Rhytidiadelphus squarrosus ; growth; heavy metal effects; cadmium; copper; lead; zinc; bioassay     

Prevalence of exposure of heavy metals and their impact on health consequences

Even in the current era of growing technology, the concentration of heavy metals present in drinking water is still not within the recommended limits as set by the regulatory authorities in different countries of the world. Drinking water contaminated with heavy metals namely; arsenic, cadmium, nickel, mercury, chromium, zinc, and lead is becoming a major health concern for public and health care professionals. Occupational exposure to heavy metals is known to occur by the utilization of these metals in various industrial processes and/or contents including color pigments and alloys. However, the predominant source resulting in measurable human exposure to heavy metals is the consumption of contaminated drinking water and the resulting health issues may include cardiovascular disorders, neuronal damage, renal injuries, and risk of cancer and diabetes. The general mechanism involved in heavy metal‐induced toxicity is recognized to be the production of reactive oxygen species resulting oxidative damage and health related adverse effects. Thus utilization of heavy metal‐contaminated water is resulting in high morbidity and mortality rates all over the world. Thereby, feeling the need to raise the concerns about contribution of different heavy metals in various health related issues, this article has discussed the global contamination of drinking water with heavy metals to assess the health hazards associated with consumption of heavy metal‐contaminated water. A relationship between exposure limits and ultimate responses produced as well as the major organs affected have been reviewed. Acute and chronic poisoning symptoms and mechanisms responsible for such toxicities have also been discussed.     

重金属对昆虫的生态生理效应

本文综述了重金属对昆虫生态生理学研究的最新进展,指出了研究的不足和应着重关注的研究方向。短期重金属暴露对昆虫有急性毒性,而长期暴露有引起昆虫对重金属污染产生适应性进化的风险。重金属对昆虫的毒性依重金属浓度、暴露时间和染毒方式而异,也会通过食物链传递和积累而影响昆虫及其天敌之间的关系。重金属对昆虫的生理毒性包括降低血细胞或血淋巴内的能量物质、引起氧化还原平衡失调、抑制细胞免疫和体液免疫、破坏昆虫细胞或组织的完整性。昆虫对重金属胁迫的生理和生态适应包括对重金属的储存和排出,解毒相关蛋白的诱导,甚至重金属耐性的进化。     

The effect of industrial pollution on Zinc,Cadmium and Copper concentration in the xylem rings of Scot's pine (Pinus sylvestris L.) and in the soil

Summary The concentrations of heavy metals, including zinc, cadmium, and copper were studied in the xylem rings of Pinus sylvestris trees and in the soil of three heavily polluted sites and one non-polluted site. There was a high correlation between the concentration of heavy metals in the soil and recently produced xylem rings. However, the initiation of industrial pollution did not abruptly increase heavy metal concentrations in the xylem rings. There was a high correlation between the zinc content of trees growing near a zinc smelter and the values obtained by other investigators for pine trees growing on the same site.     

Localization of metallothionein in the brain of rat and mouse.

Metallothionein (MT) is a low molecular mass protein inducible by heavy metals such as cadmium (Cd), zinc, and copper, and having high affinity for these metals. In the present study, we investigated the immunohistological localization of MT in the brains of rats and mice. In adult rat brain, almost no MT immunostaining was observed, whereas in adult mouse brain strong MT immunostaining was found in the ependymal cells, some glial cells, arachnoid, and pia mater. No immunostaining was detected in neurons and endothelial cells. In younger rats (1-3 weeks old), strong MT immunostaining was observed in ependymal cells, choroid plexus epithelium, arachnoid, and pia mater. The overall MT concentration in adult mouse brain appeared higher than that of the brains of young and adult rats. When adult rats were administered Cd, MT was induced not only in some glial cells, ependymal cells, arachnoid, and pia mater but also in endothelial cells. Although Cd treatment resulted in an increase in the MT immunostaining in the specific cells described above, the MT induction was not great enough to significantly affect the overall MT level in the brain. The present result suggest a possible link of MT with cell growth of choroid plexus epithelium and ependymal cells, as well as a detoxifying role of MT in the blood-brain barrier and the cerebrospinal fluid-brain barrier.     

Application of in situ cage cultures of phytoplankton for monitoring heavy metal pollution in two Norwegian fjords

A simple apparatus for in situ use of the cage culture technique for growing planktonic algae has been developed and used to follow growth and metal uptake of three diatoms in two Norwegian fjords polluted by heavy metals. Continuous pumping of sea water through a chelating resin and continuous water sampling was used to obtain average values for dissolved and particulate heavy metal content at the various test sites. The metals investigated (zinc, copper, lead, cadmium, and mercury) showed differences in the proportion of dissolved to particulate fractions.The three species of diatoms tested gave systematic growth responses to heavy metal pollution in the moderately polluted fjord; one alga died, one showed reduced growth rate at the more contaminated site compared with those at less polluted sites, while the most tolerant alga was apparently not affected. In the heavily polluted fjord only the most tolerant alga survived, showing decreasing growth rate with increasing pollution.Uptake of heavy metals increased generally with increasing heavy metal content in the sea water. The contents in the algae grown in the most polluted fjord were much higher than those obtained in the less polluted fjord, which were higher than the contents reported for algae from non-contaminated areas.     

Effect of the Heavy Metals on Developmental Stages of Ovule,Pollen, and Root Proteins in <Emphasis Type="Italic">Reseda lutea</Emphasis> L. (Resedaceae)

Heavy metals are some of the most important environmental pollutants. Excessive amounts of heavy metals adversely affect plant growth and development. Also, the presence of elevated levels of heavy metal ions triggers a wide range of cellular responses including changes in gene expression and synthesis of metal-detoxifying peptides. The overall objective of this research was to elucidate some microscopic effects of heavy metals on the formation, development, and structure of pollen, ovule, and embryo and also root proteins in Reseda lutea L. For this purpose, the vicinity of Ahangaran lead–zinc mine (Hamedan, Iran) was chosen as a polluted area where amount of some heavy metals was several times higher than the natural soils. Flowers and young buds were collected from non-polluted and polluted plants, fixed in FAA₇₀, and studied during developmental stages by light microscopy. The results showed that heavy metals can cause some abnormalities during the pollen and ovule developmental process. The number of pollen grains was decreased, and their shape was changed. Increasing in thickness of the callosic wall and stabilizing of tapetum layer were observed in polluted plants. Asymmetrical formation of ovular integuments, degradation of egg apparatus, irregular formation of embryo sac, considerable vacuolation of embryonic cells, and degeneration of embryo in the late stage of heart-shaped embryo are the results of heavy metal pollution. For protein studies, young roots were harvested from plants exposed to pollution and non-exposed to pollution at the same time. Root proteins were extracted and studied by electrophoresis. The results revealed that some new proteins were synthesized in polluted samples that probably elevate plant tolerance to heavy metals.     

重金属诱导拟南芥原生质体DNA损伤的单细胞凝胶电泳检测

以拟南芥原生质体为实验体系,研究不同浓度的3种重金属离子对拟南芥原生质体的毒性和DNA损伤的差异。结果表明,用1-5mmol·L^-1的Zn^2＋、Cd^2＋和Cu^2＋分别处理的拟南芥原生质体,2小时内活力逐渐下降,并表现出明显的浓度依赖性：与相同浓度的Cd^2＋和Cu^2＋相比,Zn^2＋对拟南芥原生质体活力的影响程度较小,表现出较低的毒性。单细胞凝胶电泳检测发现,用0．1-0．8mmol·L^-1的Zn^2＋、Cd^2＋和Cu^2＋分别处理拟南芥原生质体30分钟,以OTM值表示的原生质体DNA损伤量随重金属离子浓度的增加而递增：相同浓度（0．5mmol·L^-1）的3种重金属离子相比,Zn^2＋对原生质体的遗传毒性明显低于Cu^2＋和Cd^2＋。综合原生质体活力和DNA损伤的单细胞凝胶电泳检测结果,发现ZnO^2＋对拟南芥原生质体的遗传毒性较低,而CdO^2＋和Cu^2＋的遗传毒性较高。本研究建立的拟南芥原生质体实验体系,结合运用单细胞凝胶电泳技术,能够快速、灵敏地检测重金属对植物细胞的遗传毒性。     

The effect of in utero hypoxia on fetal heart and brain trace metals.

This study determined the effect of in utero hypoxia on fetal heart and brain pro- and antioxidant trace metals. Dunkin-Hartley guinea pigs (50–60 days gestation) were exposed to 1 h hypoxia (7% O₂/93% N₂) followed by 4 h reoxygenation in room air. Fetal hearts and brains were harvested and analyzed for copper, iron, magnesium and zinc. Fetal brain iron was significantly increased 28% after hypoxia and 35% by 1 h posthypoxia. Fetal brain magnesium demonstrated progressive decreases of 18% by 4 h posthypoxia. No significant effects of hypoxia were observed on heart trace metals. These results indicate that prooxidant metals may be increased and antioxidant metals may be decreased in posthypoxic fetal brain during a time when these tissues may be vulnerable to oxidative injury.     

Cadmium, copper and zinc in tissues of deceased copper smelter workers

Workers at a copper and lead smelter in northern Sweden have a multifactorial exposure to a number of heavy metals. The concentrations of cadmium, copper and zinc in liver, lung, kidney and brain tissues have been determined by atomic absorption spectrometry in 32 deceased long-term exposed male lead smelter workers, and compared with those of 10 male controls. Furthermore, copper and zinc levels in hair and nails were determined by energy-dispersive X-ray fluorescence.

The highest cadmium concentrations among both workers and controls were observed in kidney, followed in order by liver, lung and brain. The levels in kidney, liver and lung were all significantly higher in the workers than in the controls (p < 0.03). Among the workers relatively strong positive correlations (p < 0.03) were observed between cadmium concentrations in liver and lung, liver and kidney, liver and brain, and lung and brain. In the exposed workers a positive correlation was observed between cadmium and zinc concentrations in the kidney (r_s = 0.38; p = 0.034). This is probably mainly due to the protein metallothionein, which is stored in the kidney, binding equimolar amounts of these two metals.

The highest concentrations of copper were found in hair and nails among both workers and controls, followed in order by liver, brain, kidney and lung. The tissue concentrations of copper in brain, lung and kidney were all significantly higher among the smelter workers than in the controls (p ≤ 0.036). Copper levels in lung and age at time of death were positively correlated among the exposed workers (r_s = 0.39; p = 0.029). In the same group, positive correlations between copper and zinc concentrations in kidney (r_s = 0.45; p = 0.009) and nails (r_s = 0.68; p < 0.001) were also observed, reflecting possible biological interactions between these two metals.

Among both workers and controls, the highest zinc concentrations were found in hair, followed in order by nails, liver, kidney, brain and lung. Significantly higher tissue concentrations among the workers as compared with the reference group were noted in kidney, liver and brain (p ≤ 0.033).

Neither copper nor zinc concentrations in hair and nails seemed to provide a useful measure of the trace element status of the smelter workers.     

Influence of heavy metals on the accumulation of trimethylglycine, putrescine and spermine in food plants

Summary. Increased contents of cadmium (Cd), lead (Pb), zinc (Zn) and other heavy metals in barley plants enhanced the accumulation of trimethylglycine (betaine), putrescine and spermine. Higher contents of heavy metals in barley were caused by soil enrichment with heavy metals and by soil salinity. The highest accumulation of spermine and betaine (increase 3-fold or 5-fold in comparison to untreated soil substrates) was obtained at the highest concentration of heavy metals in plants. Consequently the betaine-N / protein-N-ratio and the spermine-N / protein-N-quotient increased 3-fold in plants with high heavy metal contents. The biomass formation was not changed significantly by the different experimental treatments. Received January 28, 2000 Accepted March 1, 2000