Modulation of nutrient acquisition and polyamine pool in salt-stressed wheat (Triticum aestivum L.) plants inoculated with arbuscular mycorrhizal fungi

Two wheat (Triticum aestivum L.) cultivars, Sids 1 and Giza 168, were grown under non-saline or saline conditions (4.7 and 9.4 dS m^?1) with and without arbuscular mycorrhizal fungi (AMF) inoculation. Salt stress considerably decreased root colonization, plant productivity and N, P, K⁺, Fe, Zn and Cu concentrations, while it increased Na⁺ level, particularly in Giza 168. Mycorrhizal colonization significantly enhanced plant productivity and N, P, K⁺, Fe, Zn and Cu acquisition, while it diminished Na⁺ uptake, especially in Sids 1. Salinity increased putrescine level in Giza 168, however, values of spermidine and spermine increased in Sids 1 and decreased in Giza 168. Mycorrhization changed the polyamine balance under saline conditions, an increase in putrescine level associated with low contents of spermidine and spermine in Giza 168 was observed, while Sids 1 showed a decrease in putrescine and high increase in spermidine and spermine. Moreover, mycorrhizal inoculation significantly reduced the activities of diamine oxidase and polyamine oxidase in salt-stressed wheat plants. Modulation of nutrient acquisition and polyamine pool can be one of the mechanisms used by AMF to improve wheat adaptation to saline soils. This is the first report dealing with mycorrhization effect on diamine oxidase and polyamine oxidase activities under salt stress.     

Effects of sub-chronic aluminum chloride exposure on rat ovaries

Aims

This experiment investigated the effects of sub-chronic aluminum chloride (AlCl₃) exposure on rat ovaries.

Main methods

Eighty female Wistar (5 weeks old) rats, weighed 110–120 g, were randomly divided into four treatment groups: control group (CG), low-dose group (LG, 64 mg/kg BW AlCl₃), mid-dose group (MG, 128 mg/kg BW AlCl₃) and high-dose group (HG, 256 mg/kg BW AlCl₃). The AlCl₃ was administered in drinking water for 120 days. The ovarian ultrastructure was observed. The activities of acid phosphatase (ACP), alkaline phosphatase (ALP), succinate dehydrogenase (SDH), Na⁺–K⁺-ATPase, Mg^2 +-ATPase and Ca^2 +-ATPase, the contents of Fe, Cu and Zn, and the protein expression of follicle-stimulating hormone receptor (FSHR) and luteinizing hormone receptor (LHR) in the ovary were determined.

Key findings

The results showed that the structure of the ovary was disrupted, the activities of ALP, ACP, SDH, Na⁺–K⁺-ATPase, Mg^2 +-ATPase and Ca^2 +-ATPase, the contents of Zn, Fe and the protein expression of FSHR and LHR were lowered, and the content of Cu was increased in AlCl₃-treated rats than those in control.

Significance

The results indicate that sub-chronic AlCl₃ exposure caused the damage of the ovarian structure, the disturbed metabolism of Fe, Zn and Cu and the decreased activities of Na⁺–K⁺-ATPase, Mg^2 +-ATPase and Ca^2 +-ATPase in the ovary, which could result in suppressed energy supply in the ovary. A combination of suppression of energy supply and reduction of expression of FSHR and LHR could inhibit ovulation and corpus luteum development, leading to infertility in female rats.     

The Effect of Electrolyte Type and Concentration on the Release of Cd,Cu, Ni,and Zn in Some Contaminated Calcareous Soils

This paper reports the results of desorption experiments of cadmium (Cd), copper (Cu), nickel (Ni), and zinc (Zn) from some contaminated calcareous soils under four electrolyte types (CaCl₂, MgCl₂, NaCl and Na₂SO₄) with different electrolyte concentrations (0.5, 4 and 10 mM). Among electrolytes, CaCl₂ significantly released more metals from soils. There was a negative relationship between total Cu and Zn content and percentage of Cu and Zn released (average of electrolyte concentrations) using CaCl₂ solution, indicating a higher Cu and Zn released when their total content was low. Generally, Cd, Cu, and Zn speciation was affected by both type of electrolytes and their concentrations, whereas Ni speciation stayed mostly stable and was almost unaffected by applied solutions. It can be suggested that beside competition with cations, chloro-complexation is important parameter in Cd release, while CuOH⁺, and to some extent ZnOH⁺ are important species affecting release of Cu and Zn. The distribution coefficient (K_d) values for each metals greatly varied with the types of electrolytes and electrolyte concentration. On the basis of average percentage of metal released under different electrolytes and concentrations the following sequences was found: Cd > Cu > Ni > Zn. The results are important in understanding the mobility of metals under different solutions and indicating that, Cd and Zn soils may pose a higher and lower mobility and ecological risk in contaminated calcareous soils, respectively.     

Distribution of manganese and other biometals in <Emphasis Type="Italic">flatiron</Emphasis> mice

Flatiron (ffe) mice display features of “ferroportin disease” or Type IV hereditary hemochromatosis. While it is known that both Fe and Mn metabolism are impaired in flatiron mice, the effects of ferroportin (Fpn) deficiency on physiological distribution of these and other biometals is unknown. We hypothesized that Fe, Mn, Zn and/or Cu distribution would be altered in ffe/+ compared to wild-type (+/+) mice. ICP-MS analysis showed that Mn, Zn and Cu levels were significantly reduced in femurs from ffe/+ mice. Bone deposits reflect metal accumulation, therefore these data indicate that Mn, Zn and Cu metabolism are affected by Fpn deficiency. The observations that muscle Cu, lung Mn, and kidney Cu and Zn levels were reduced in ffe/+ mice support the idea that metal metabolism is impaired. While all four biometals appeared to accumulate in brains of flatiron mice, significant gender effects were observed for Mn and Zn levels in male ffe/+ mice. Metals were higher in olfactory bulbs of ffe/+ mice regardless of gender. To further study brain metal distribution, ⁵⁴MnCl₂ was administered by intravenous injection and total brain ⁵⁴Mn was measured over time. At 72 h, ⁵⁴Mn was significantly greater in brains of ffe/+ mice compared to +/+ mice while blood ⁵⁴Mn was cleared to the same levels by 24 h. Taken together, these results indicate that Fpn deficiency decreases Mn trafficking out of the brain, alters body Fe, Mn, Zn and Cu levels, and promotes metal accumulation in olfactory bulbs.     

Kinetics and thermodynamics of metal‐binding to histone deacetylase 8

Histone deacetylase 8 (HDAC8) was originally classified as a Zn(II)-dependent deacetylase on the basis of Zn(II)-dependent HDAC8 activity in vitro and illumination of a Zn(II) bound to the active site. However, in vitro measurements demonstrated that HDAC8 has higher activity with a bound Fe(II) than Zn(II), although Fe(II)-HDAC8 rapidly loses activity under aerobic conditions. These data suggest that in the cell HDAC8 could be activated by either Zn(II) or Fe(II). Here we detail the kinetics, thermodynamics, and selectivity of Zn(II) and Fe(II) binding to HDAC8. To this end, we have developed a fluorescence anisotropy assay using fluorescein-labeled suberoylanilide hydroxamic acid (fl-SAHA). fl-SAHA binds specifically to metal-bound HDAC8 with affinities comparable to SAHA. To measure the metal affinity of HDAC, metal binding was coupled to fl-SAHA and assayed from the observed change in anisotropy. The metal K_D values for HDAC8 are significantly different, ranging from picomolar to micromolar for Zn(II) and Fe(II), respectively. Unexpectedly, the Fe(II) and Zn(II) dissociation rate constants from HDAC8 are comparable, k_off ∼0.0006 s⁻¹, suggesting that the apparent association rate constant for Fe(II) is slow (∼3 × 10³ M⁻¹ s⁻¹). Furthermore, monovalent cations (K⁺ or Na⁺) that bind to HDAC8 decrease the dissociation rate constant of Zn(II) by ≥100-fold for K⁺ and ≥10-fold for Na⁺, suggesting a possible mechanism for regulating metal exchange in vivo. The HDAC8 metal affinities are comparable to the readily exchangeable Zn(II) and Fe(II) concentrations in cells, consistent with either or both metal cofactors activating HDAC8.     

Effects of soil acidification and subsequent leaching on levels of extractable nutrients in a soil

Summary The effects of soil acidification (pH values from 6.5 to 3.8), and subsequent leaching, on levels of extractable nutrients in a soil were studied in a laboratory experiment. Below pH 5.5, acidification resulted in large increases in the amounts of exchangeable Al in the soil. Simultaneously, exchangeable cations were displayed from exchange sites and Ca, Mg, K and Na in soil solution increased markedly. With increasing soil acidification, increasing amounts of cations were leached; the magnitude of leaching loss was in the same order as the cations were present in the soil: Ca²⁺>Mg²⁺>K⁺>Na⁺. Soil acidification appeared to inhibit nitrification since in the unleached soils, levels of NO ₃ ⁻ clearly declined below pH 5.5 and at the same time levels of NH ₄ ⁺ increased greatly. Significant amounts of NH ₄ ⁺ and larger amounts of NO ₃ ⁻ , were removed from the soil during leaching. Concentrations of NaHCO₃-extractable phosphate remained unchanged between pH 4.3 and 6.0 but were raised at higher and lower pH values. No leaching losses of phosphate were detected. For the unleached soils, levels of EDTA-extractable Mn and Zn increased as the soil was acidified whilst levels of extractable Fe were first decreased and then increased greatly and those for Cu were decreased slightly between pH 6.5 and 6.0 and then unaffected by further acidification. Significant leaching losses of Mn and Zn were observed at pH values below 5.5 but losses of Fe were very small and those of Cu were not detectable.     

Mechanisms of dietary Cu uptake in freshwater rainbow trout: evidence for Na-assisted Cu transport and a specific metal carrier in the intestine

Copper (Cu) is both a vital nutrient and a potent toxicant. The objective of this study was to analyze the mechanistic nature of intestinal Cu transport in rainbow trout using radiolabeled Cu (⁶⁴Cu) and an in vitro gut sac technique. Reduction of mucosal NaCl levels inhibited Cu transport while increase caused stimulation; Na₂SO₄ had an identical effect, implicating Na⁺ rather than the anion. These responses were unrelated to solvent drag, osmotic pressure or changes in transepithelial potential. The presence of elevated luminal Ag stimulated Cu and Na⁺ uptake. Phenamil caused a partial inhibition of both Cu and Na⁺ uptake while hypercapnia stimulated Na⁺ and Cu transport. Cu uptake was sensitive to luminal pH and inhibited by a tenfold excess of Fe and Zn. These factors had no effect on Na⁺ uptake. On the basis of these results we propose a novel Na⁺-assisted mechanism of Cu uptake wherein the Na⁺ gradient stimulates an increase in the H⁺ concentration of the brushborder creating a suitable microenvironment for the effective transport of Cu via either DMT1 or Ctr1.     

Mechanistic characterization of gastric copper transport in rainbow trout

An in vitro gut-sac technique and ⁶⁴Cu as a radiotracer were used to characterize gastric copper (Cu) transport. Cu transport was stimulated by low luminal pH (4.0 vs. 7.4), to a greater extent than explained by the increased availability of the free Cu²⁺ ion. At pH = 4.0, uptake kinetics were indicative of a low affinity (K _m = 525 μmol L⁻¹), saturable carrier-mediated component superimposed on a large linear (diffusive and/or convective) component, with about 50% occurring by each pathway at Cu = 50 μmol L⁻¹. Osmotic gradient experiments showed that solvent drag via fluid transport may play a role in Cu uptake via the stomach, in contrast to the intestine. Also unlike the intestine, neither the Na⁺ gradient, high Ag, nor phenamil had any influence on gastric Cu transport, and a tenfold excess of Fe and Zn failed to inhibit Cu uptake. These findings indicate that neither Na⁺-dependent pathways nor DMT1 are likely candidates for carrier-mediated Cu transport in the stomach. We have cloned a partial cDNA sequence for the copper transporter Ctr1, and show its mRNA expression in all segments of the trout gastrointestinal tract, including the stomach. Based on the fact that this transporter is functional at low pH conventionally found in the stomach lumen, we suggest Ctr1 is a pathway for gastric Cu transport in trout. Extreme hypoxia inhibited Cu uptake. High P_\textCO2 P_{{{\text{CO}}_{2} }} levels (7.5 torr) increased Cu uptake and acetazolamide (100 μmol L⁻¹) significantly inhibited Cu uptake, indicating carbonic anhydrase activity was involved in gastric Cu transport. Transport of Cu was insensitive to bafilomycin (10 μmol L⁻¹) suggesting a V-ATPase did not play a direct role in the process. Expression (mRNA) of H ⁺ , K ⁺-ATPase, carbonic anhydrase 2, and the α-3 isoform of Na ⁺–K ⁺-ATPase were observed in the stomach. We suggest these enzymes facilitate Cu transport in the stomach indirectly as part of a physiological mechanism exporting H⁺ to the cell exterior. However, pre-treatment with the H ⁺ , K ⁺-ATPase proton pump blocker omeprazole did not affect gastric Cu transport, suggesting that other mechanisms must also be involved.     

Phytotoxic Effects of Cd, Zn, Pb, Cu and Fe on Sinapis Alba L. Seedlings and their Accumulation in Roots and Shoots

The inhibitory effects of Cd, Cu, Zn, Pb, and Fe on root elongation, contents of photosynthetic pigments, and metal accumulation in the roots and shoots of Sinapis alba were assessed. On the basis of growth inhibition metals can be arranged in a order Cu > Cd > Fe = Zn > Pb. All the metals, except Fe, were accumulated in significantly higher amount in the roots than in the shoots. Cd, Zn, Cu and Pb reduced chlorophyll a, and especially chlorophyll b content, and Zn and Pb reduced the carotenoid content, but less than that of chlorophyll a+b. The plants contained the highest concentration of Cd, and the lowest concentration of Zn.     

Comparison of the nutrient ecology of coastal Banksia grandis elfinwood (windswept shrub‐like form) and low trees,Cape Leeuwin‐Naturaliste National Park,Western Australia

Abstract Trees growing along windy coasts often have canopies that are greatly reduced in size by the sculpting effects of wind and salt spray. Trees with environmentally reduced stature are called elfinwood (windswept shrub‐form or krummholz) and are ecologically important because they represent outposts growing at the limit of tree success. The purpose of this study was to assess if Banksia grandis elfinwood growing at Cape Leeuwin had a different nutrient status than normal low‐form (LF) trees growing nearby, and if nutrient deficiencies, toxicities and/or imbalances were among the limiting factors imposed on elfinwood. The concentrations of N, P, K, Ca, Mg, Na, Cl^–, Fe, Mn, Zn, Cu, Mo and B were analysed for mature green foliage, immature foliage, foliage litter, flowers and soil. When the elfinwood and LF trees were compared, the foliar nutrient status was generally similar, except that elfinwood foliage had significantly higher mean concentrations of N, Zn and Cu, while LF trees had higher Fe and Mn contents. Many nutrients were conserved before leaves were shed in both elfinwood and LF trees, including N, P, K, Na, Cl^–, Mn and Cu (LF trees also conserved Ca and Mg). However, elfinwood and LF tree‐litter contained significantly higher Fe concentrations than green foliage (elfinwood litter also had higher levels of Mg and B). It is tempting to suggest that the translocation of Fe into leaves before they were shed is a regulation mechanism to prevent Fe toxicity, or imbalance in the Fe : Mn ratio. Proteoid roots strongly acidify the soil to mobilize P, which also chemically reduces Fe⁺³ to plant‐available Fe⁺². The increased supply of Fe⁺² in the rhizosphere, caused by the action of proteoid roots, might tend to defeat self‐regulation of Fe uptake. It is possible that excess Fe accumulation in the plant might be regulated, in part, by exporting Fe into the leaves before they are shed. The nutrient status of B. grandis elfinwood is compared with mountain elfinwood of North America. The extreme habitat of coastal elfinwood provides many theoretical pathways for nutrient limitation, but B. grandis elfinwood at Cape Leeuwin does not appear to be nutrient deficient.     

Copper Uptake and Its Effect on Metal Distribution in Root Growth Zones of Commelina communis Revealed by SRXRF

To explore the copper uptake mechanisms by the Cu-tolerant plant Commelina communis, the contents of Cu and other metals (including Fe, Zn, and Mn) in roots were detected using atomic absorption spectrometer under transporter inhibitors, partial element deficiency, or Cu excess treatments, while distribution characters of Cu and other metals in root growth zones were investigated by synchrotron radiation X-ray fluorescence spectroscopy (SRXRF). Cu uptake was inhibited by the uncoupler DNP and P-type ATPase inhibitor Na₃VO₄, not by the Ca²⁺ ion channel inhibitor LaCl₃, suggesting that Cu could probably be assimilated actively by root and be related with P-type ATPase, but not through Ca²⁺ ion channel. Fe or Zn deficiency could enhance Cu uptake, while 100 μM Cu inhibited Fe, Zn, and Mn accumulation in roots significantly. Metal distribution under 100 μM Cu treatment was investigated by SRXRF. High level of Cu was found in the root meristem, and higher Cu concentrations were observed in the vascular cylinder than those in the endodermis, further demonstrating the initiative Cu transport in the root of C. communis. Under excess Cu stress, most Fe was located in the epidermis, and Fe concentrations in the endodermis were higher than those in the vascular cylinder, suggesting Cu and Fe competition not only in the epidermal cells but also for the intercellular and intracellular transport in roots. Zn was present in the meristem and the vascular cylinder similar to Cu. Cu and Zn showed a similar pattern. Mn behaves as Zn does, but not like Fe.     

Acquisition of Cu, Zn, Mn and Fe by mycorrhizal maize (Zea mays L.) grown in soil at different P and micronutrient levels

Sustainability of soil-plant systems requires, among other things, good development and function of mycorrhizal symbioses. The effects of P and micronutrient levels on development of an arbuscular mycorrhizal fungus (AMF) and uptake of Zn, Cu, Mn and Fe by maize (Zea mays L.) were studied. A pot experiment with maize either inoculated or not with Glomus intraradices was conducted in a sand:soil (3 :1) mix (pH 6.5) in a greenhouse. Our goal was to evaluate the contribution of mycorrhizae to uptake of Cu, Zn, Mn and Fe by maize as influenced by soil P and micronutrient levels. Two levels of P (10 and 40 mg kg⁻¹ soil) and three levels of a micronutrient mixture: 0, 1X and 2X (1X contained, in mg kg⁻¹ soil, 4.2 Fe, 1.2 Mn, 0.24 Zn, 0.06 Cu, 0.78 B and 0.036 Mo), were applied to pots. There were more extraradical hyphae at the low P level than at the high P level when no micronutrients were added to the soil. Root inoculation with mycorrhiza and application of micronutrients increased shoot biomass. Total Zn content in shoots was higher in mycorrhizal than non-mycorrhizal plants grown in soils with low P and low or no micronutrient addition. Total Cu content in shoots was increased by mycorrhizal colonization when no micronutrients were added. Mycorrhizal plants had lower Mn contents than non-mycorrhizal plants only at the highest soil micronutrient level. AMF increased total shoot Fe content when no micronutrients were added, but decreased shoot Fe when plants were grown at the high level of micronutrient addition. The effects of G. intraradices on Zn, Cu, Mn, and Fe uptake varied with micronutrient and P levels added to soil. Accepted: 27 December 1999     

Levels of iron, zinc, and copper in aqueous humor, lens, and serum in nondiabetic and diabetic patients

The purpose of this study was to investigate iron (Fe), zinc (Zn), and copper (Cu) levels of aqueous humor, lens, and serum in nondiabetics and diabetics and to determine the effects of diabetes on Fe, Zn, and Cu contents in the lens. Fe, Zn, and Cu contents of aqueous humor, lens, and serum samples of 19 patients (9 nondiabetic patients with a mean age of 62.3±5.4 yr, and 10 diabetic patients with a mean age of 59.5±5.9 yr) were analyzed by atomic absorption spectrometry using a prospective study design. The lens levels of Cu in diabetic patients were significantly higher compared with nondiabetic patients (p=0.02); however; there was no difference in the other elements (Zn, Fe; p=0.28, p=0.74, respectively). The levels of Fe, Zn, and Cu in the aqueous humor and serum of diabetic patients were not found to be statistically significant when compared to nondiabetics (p=0.46, p=0.11, p=0.18, and p=0.22, p=0.43, p=0.72, respectively). These results demonstrate that increased Cu content of the lens presumably has a greater association with the development of lens opacification in diabetics than Zn and Fe content.     

Changes in the total content of iron,copper, and zinc in serum,heart, liver,spleen, and skeletal muscle tissues of rats infected withTrypanosoma cruzi

The effects of theTrypanosoma cruzi infection on the total content of the essential trace elements iron (Fe), copper (Cu), and zinc (Zn) in serum, heart, liver, spleen, and skeletal muscle were determined in “Wistar” rats inoculated with reticulotropic “Y” strain trypanosoms (Tryps) in their slender blood form. The 250 rats were divided in two groups of 80 rats (L-1 and L-2) and one of 90 (C) used as controls. L-1 and L-2 were inoculated with 2×10⁵ and 5×10² Tryps, respectively. Ten rats of the C group were killed the inoculation day (i), and ten rats of each group chosen at random were killed and blood parasitemia determined at 5, 10, 15, 20, 25, 30, 60, and 90 post-i days covering the infection acute-phase myocarditis. Previously cryohomogenized and lyophilized tissues were digested in an HNO₃ ⁻ H₂O₂ mixture with the aid of a microwave oven, and the elements Fe, Cu, and Zn were determined by Flame Atomic Absorption Spectrometry. Generally, more intense changes were observed in the L-1 group. Serum Fe and Zn levels are lower and Cu levels higher in groups L-1 and L-2 than in C. However, Fe is not significantly sequestered in the liver during the acute phase of the infection as expected, but of the tissues studied, the spleen was the main site of Fe binding. Zn tended to increase in all tissues, except in the spleen, where during the acute phase of the infection, the total content of Zn in groups L-1 and L-2 was lower than in group C. Cu increased mainly in the spleen and muscle. In general, each tissue presented its own pattern of redistribution related to its nature, functions, and number of parasites inoculated, and these patterns may have been altered by the tropism of the parasite.     

微量元素五项联合红细胞四项检测在小儿缺铁性贫血中的诊断价值

目的:探讨微量元素五项联合红细胞四项检测在小儿缺铁性贫血(IDA)中的诊断价值。方法:选取2013年1月至2016年1月于我院进行治疗的小儿缺铁性贫血(IDA)患儿130例作为缺铁性贫血(IDA)组及同期于我院进行常规体检的健康儿童40例作为对照组。检测和比较其钙、镁、铜、铁、锌、血红蛋白(Hb)、平均红细胞体积(MCV)、平均血红蛋白含量(MCH)、平均血红蛋白浓度(MCHC)水平,并比较微量元素五项、红细胞四项检查以及两者联合对IDA患儿的诊断效能。结果:缺铁性贫血(IDA)组患儿微量元素镁、铁、锌含量以及Hb、MCV、MCH及MCHC水平均明显低于对照组儿童,差异具有统计学意义(P0.05);两组儿童微量元素钙、铜含量比较差异不显著(P0.05)。微量元素五项联合红细胞四项的灵敏度、特异度、阳性预测值及阴性预测值均明显高于微量元素五项和红细胞四项(P0.05)。结论:IDA患儿微量元素镁、铁、锌含量以及Hb、MCV、MCH及MCHC水平较低,微量元素五项联合红细胞四项检测可以提高小儿IDA的检出率。     

Room temperature solvent extraction for simple and fast determination of total concentration of Ca,Cu, Fe,Mg, Mn,and Zn in bee pollen by FAAS along with assessment of the bioaccessible fraction of these elements using in vitro gastrointestinal digestion

Background and aimBee pollen is recognized to be a source of different nutrients, including minerals. As a food supplement, its quality and safety due to concentrations of essential macro- and microelements, and harmful trace elements has to be verified. Fast and simple element analysis of bee-collected pollen can be regarded as an important part of its quality assurance and control. The present study aimed at developping a new method for determination of selected elements (Ca, Cu, Fe, Mg, Mn, Zn) of bee pollen based on solvent extraction and completely avoiding a high temperature treatment with concentrated reagents. In addition, in vitro gastrointestinal digestion was used to assess bioavailability of elements from this food supplement.MethodsBee pollen samples were dried and pulverized. Total concentrations of Ca, Cu, Fe, Mg, Mn, and Zn were determined by flame atomic absorption spectrometry (FAAS) in sample solutions obtained by wet digestion (WD) in concentrated HNO₃ or alternatively by solvent extraction (SE) with diluted solutions of HNO₃. Gastrointestinal digestion was mimicked using simulated solutions of gastric and intestinal juices followed by determination of Ca, Cu, Fe, Mg, Mn and Zn concentrations in the bioaccessible fraction by FAAS.ResultsA new simple and fast method for determination of total concentrations of Ca, Cu, Fe, Mg, Mn, and Zn in bee pollen was developed and validated. The method combined room temperature, two-hour SE with 0.5 mol L⁻¹ HNO₃ with FAAS measurements versus simple standard solutions. It provided precision within 1–5 % and trueness better than 8%, and was shown to be suitable for fast analysis of different polyfloral bee pollens. In vitro gastrointestinal digestion revealed that elements were well (70–85 % for Ca, Mg) and fairly (27–43 % for Cu, Fe, Mn, and Zn) bioaccessible from bee pollen. By pouring with water and swelling overnight, bioaccessibility of studied elements from such prepared bee pollen was increased on average by less than 15 % (Mn), 20 % (Ca, Cu, Fe, Zn) or 30 % (Mn).ConclusionsAvoiding long-lasting, high-temperature wet digestion with concentrated reagents, the proposed sample treatment along with FAAS provided precise and true results of total concentrations of Ca, Cu, Fe, Mg, Mn, and Zn in bee pollen. The method was simple and fast, and enabled to analyze a higher number of samples. Simulated gastrointestinal digestion of bee pollen have shown for the first time that Ca and Mg are the most bioaccessible from this bee product. Bioaccessibility of Cu, Fe, Mg, and Zn from bee pollen are close to or lower than 40 %.     

Some Physiological Aspects of Copper and Zinc Tolerance in Agrostis tenuis Sibth.: Cell Elongation and Membrane Damage

Three clones of Agrostis tenuis Sibth. were studied with respectto the effects of Zn and Cu on the growth of root segments excisedfrom the zone of cell elongation. Elongation growth in segmentsfrom a Cu-tolerant and a Zn-tolerant clone was inhibited toa lesser extent by Cu and Zn respectively than was the growthof a clone which was not tolerant to these metals. Concentrationsof Cu²⁺ which inhibited root growth also caused leakage of K⁺from the cells but toxic concentrations of Zn²⁺ did not induceK⁺ leakage. Copper induced a higher rate of K⁺ leakage at 25than at 0 °C. The impllcations of these results for thesite of the toxic effects of Zn and Cu and the nature of theresistance mechanisms are discussed.     

The effects of copper, iron and zinc on digestive enzyme activity in the hybrid tilapia Oreochromis niloticus (L.) ×Oreochromis aureus (Steindachner)

The present experiment was conducted to study effects of Cu, Fe and Zn on activities of digestive enzymes of the hybrid tilapia Oreochromis niloticus×Oreochromis aureus. The acidic protease activities increased 65·5 and 55·1% by addition of homogenates of digesta‐containing stomach with copper (75 mg l⁻¹) and zinc (50 mg l⁻¹) respectively. Addition of Cu and Zn increased the activities of protease in the hepatopancreas homogenates by 132·7 and 38·1% respectively, and reduced the activity of protease in the digesta‐containing intestine homogenates by 11·0 and 13·8% respectively. Addition of Fe (50 mg l⁻¹) increased the acidic protease activity by 96·7% but did not alter the activities of protease in the intestine and hepatopancreas. Addition of Cu markedly inhibited activities of amylase in intestine and hepatopancreas homogenates, while Zn addition showed no effects. Addition of Fe reduced activities of amylase in the intestine homogenates by 47·9% but had no effect on amylase activities in the hepatopancreas. When Cu (75 mg kg⁻¹), Fe (50 mg kg⁻¹) and Zn (50 mg kg⁻¹) were supplemented to basal diet for 3 weeks, the activities of amylase in hepatopancreas homogenates increased 125·3, 215·6 and 70·0%, respectively, the activities of amylase in intestine increased 79·8, 74·6 and 48·5%, respectively, and the activities of lipase in intestine increased 90·5, 149·8 and 84·0%, respectively. Supplementation of Cu, Fe or Zn into diet had no effects on activity of protease in all digestive organs. Therefore, the results suggest that effects of Cu, Fe and Zn on activity of digestive enzymes in vitro were different from those seen in vivo, and that the positive effects of Cu, Fe and Zn supplemented to fish diet would be valuable information for formulating fish feed.     

Changes in the selected hematological parameters and gill Na/K ATPase activity of juvenile crucian carp Carassius auratus during elevated ammonia exposure and the post-exposure recovery

The increase in concentration of ammonia in lake water during the degradation of algal blooms may last for several weeks and thus cause chronic toxicity to aquatic organisms. The purpose of this study was to assess the chronic toxicity of ammonia on the selected hematological parameters and gill Na⁺/K⁺ ATPase activity of juvenile crucian carp Carassius auratus during elevated ammonia exposure and the post-exposure recovery. Juvenile crucian carp were exposed in different ammonia solutions for 45 days and then immediately transferred to pristine freshwater to initiate a 15-day recovery period. Results showed sub-lethal ammonia significantly deters growth and a 15-day recovery period was not sufficient for the fish to compensate for the loss of growth. The fish exhibited a continuous decrease in red blood cell (RBC), the total hemoglobin (Hb), and gill Na⁺/K⁺ ATPase activity as the concentration of NH₃-N increased. After the 15-day recovery period, RBC, Hb, and gill Na⁺/K⁺ ATPase activity had recovered to similar levels as the controls.     

Leaf cell membrane stability‐based mechanisms of zinc nutrition in mitigating salinity stress in rice

• Excess salt affects about 955 million ha of arable land worldwide, and 49% of agricultural land is Zn‐deficient. Soil salinity and zinc deficiency can intensify plant abiotic stress. The mechanisms by which Zn can mitigate salinity effects on plant functions are not well understood.
• We conducted an experiment to determine how Zn and salinity effects on rice plant retention of Zn, K⁺ and the salt ion Na⁺ affect chlorophyll formation, leaf cell membrane stability and grain yield. We examined the mechanisms of Zn nutrition in mitigating salinity stress by examining plant physiology and nutrition. We used native Zn‐deficient soils (control), four salinity (EC ) and Zn treatments – Zn 10 mg·kg^?1 (Zn₁₀), EC 5 dS ·m^?1 (EC ₅), Zn₁₀+EC ₅ and Zn₁₅+EC ₅, a coarse rice (KS ‐282) and a fine rice (Basmati‐515) in the study.
• Our results showed that Zn alone (Zn₁₀) significantly increased rice tolerance to salinity stress by promoting Zn/K⁺ retention, inhibiting plant Na⁺ uptake and enhancing leaf cell membrane stability and chlorophyll formation in both rice cultivars in native alkaline, Zn‐deficient soils (P  <  0.05). Further, under the salinity treatment (EC ₅), Zn inputs (10–15 mg·kg^?1) could also significantly promote rice plant Zn/K⁺ retention and reduce plant Na⁺ uptake, and thus increased leaf cell membrane stability and grain yield. Coarse rice was more salinity‐tolerant than fine rice, having significantly higher Zn/K⁺ nutrient retention.
• The mechanistic basis of Zn nutrition in mitigating salinity impacts was through promoting plant Zn/K⁺ uptake and inhibiting plant Na⁺ uptake, which could result in increased plant physiological vigour, leaf cell membrane stability and rice productivity.