Effect of algal growth on the availability of phosphorus,iron and manganese in rice soils

Summary Laboratory experiments were conducted to study the effect of algal growth on the change of (I) pH, (II) available phosphorus and (III) solubility of iron and manganese content in five waterlogged alluvial rice soils of West Bengal, India. The results showed that the algal growth initially caused an increase in the soil pH, which later declined to the original value in some of the soils. The available phosphorus content decreased upto 90 days of their growth and began to increase towards the later period of incubation. The drastic fall of water soluble plus exchaneable manganese content of the soils due to algal growth was accompanied by similar increase in reducible manganese content. No appreciable change in water soluble plus exchangeable ferrous iron content was encountered but theN-NH₄OAC(pH 3) extractable iron due to algal growth progressively decreased with the progress of the incubation period.     

Groundwater input affecting plant distribution by controlling ammonium and iron availability

Question: How does groundwater input affect plant distribution in Alnus glutinosa (black alder) carrs? Location: Alder carrs along the river Meuse, SE Netherlands. Methods: Three types of site, characterized by groundwater flow, were sampled in 17 A. glutinosa carrs. Vegetation and abiotic data (soil and water chemistry) were collected and analysed using a Canonical Correspondence Analysis. Based on the results, a laboratory experiment tested the effect of groundwater input (Ca²⁺) on pore water chemistry (NH₄⁺ availability). Results: Environmental factors indicating groundwater input (Ca²⁺ and Fe²⁺), correlating with the NH⁺₄ concentration in the pore water, best explained the variation in plant distribution. NH₄⁺ availability was determined by Ca²⁺ input via the groundwater and subsequent competition for exchange sites in the sediment. As a result, nutrient‐poor seepage locations fully fed by groundwater were dominated by small iron resistant plants such as Caltha palustris and Equisetum fluviatile. More nutrient‐rich locations, fed by a combination of groundwater and surface water, allowed the growth of taller iron resistant plant species such as Carex paniculata. Nutrient‐rich locations with stagnating surface water were hardly fed by groundwater, allowing the occurrence of fast growing and less iron tolerant wetland grasses such as Glyceria fluitans and G. maxima. Conclusion: Groundwater input affects plant composition in A. glutinosa carrs along the river Meuse by determining nutrient availability (ammonium) and concentrations of toxic iron.     

Groundwater-induced redox-gradients control soil properties and phosphorus availability across four headwater wetlands, New York, USA

Hydrochemical patterns across groundwater-fed wetlands, especially carbonate and redox gradients, can influence phosphorus (P) availability by controlling its distribution among different soil pools. We explored these linkages by comparing shallow (5–20 cm) soil properties along groundwater flowpaths in two rich fens, a marl fen, and a poor fen. Organic matter content, bulk density, and total elemental content varied more with depth to underlying drift materials than with water table fluctuation, but also were influenced by groundwater discharge, which stabilized water table elevations and controlled redox conditions. Total sulfur and calcium content increased where pore-water chemistry indicated active iron and sulfate reduction. Calcium mineral dynamics, however, did not appear to influence P availability: first, calcium carbonate (CaCO₃) accounted for <2% of the soil composition, except in the marl fen where it accounted for 20–25% of the soil composition. Second, Ca-bound P pools, determined from hydrochloric extraction of wet soil samples, accounted for <25% of the inorganic soil P pool. In contrast, iron-bound P determined from bicarbonate-buffered dithionite solution, accounted for 50–80% of the inorganic soil P, and increased where there was evidence of groundwater mixing, as did P release rates inferred from incubated anion resin bags. The total carbon and phosphorus content of organic-rich soils as well as available and labile P pools were strongly correlated with pore-water iron and alkalinity concentrations. Groundwater discharge and resulting hydrochemical gradients explained significant variation in soil composition and P availability across each site. Results highlight the importance of conducting biogeochemical studies in the context of a site’s shallow geologic setting and suggest mechanisms supporting the diverse plant species unique to groundwater wetlands.     

Effect of salinity,alkalinity and iron sources on availability of iron

The availability of iron from added iron sources was studied in normal and salt affected soils in the laboratory. All forms of iron decreased with increase in salinity and alkalinity, the lowest amount being recorded in 8 E.Ce+40 ESP soil. All the forms of iron in all the soils decreased gradually with increase in incubation period. Addition of iron in organic and inorganic forms increased all the forms of iron in all the soils. In general, iron Ke-Min and Rayplex were better in keeping higher iron availability for longer time.     

饲料钙磷含量和钙磷比对生长期大鼠体重增长与骨骼发育的影响

目的研究饲料中不同的钙磷含量和钙磷比对生长期实验大鼠体重及骨骼发育的影响。方法采用完全随机化设计方案,取1月龄（100±10）g清洁级Wistar雄性大鼠70只,随机分成7组,每组10只,饲喂7种不同钙磷含量的饲料。自由采食,饮用去离子水,试验期42 d。各组于实验开始和结束时称重。试验结束后,处死动物并分离一侧胫骨和股骨用于实验指标的检测。结果当保持饲料中钙或者磷中的一种含量不变的情况下,调整另一种的含量即改变钙磷比例时,1.2∶1的正常钙磷比例组生长期实验大鼠增重及骨骼发育要明显好于0.4∶1的低钙磷比例组和4∶1的高钙磷比例组（P〈0.05）,并且当保持饲料中钙磷比例不变时,即使在低钙低磷和高钙高磷进食水平下,正常钙磷比例组大鼠增重及骨骼发育都较好,但低钙磷比例组和高钙磷比例组大鼠增重及骨骼发育则由于饲料中钙磷含量的变化波动较大（P〈0.05）。结论饲料中钙和磷需按比例添加,如果两者绝对含量相差过大就会影响到动物的增重及骨骼发育,本实验证明饲料钙磷比为1.2∶1时生长期实验大鼠增重及骨骼发育较好。     

Differential effects of basolateral and apical iron supply on iron transport in Caco-2 cells

Iron homeostasis in the human body is maintained primarily through regulation of iron absorption in the duodenum. The liver peptide hepcidin plays a central role in this regulation. Additionally, expression and functional control of certain components of the cellular iron transport machinery can be influenced directly by the iron status of enterocytes. The significance of this modulation, relative to the effects of hepcidin, and the comparative effects of iron obtained directly from the diet and/or via the bloodstream are not clear. The studies described here were performed using Caco-2 cell monolayers as a model of intestinal epithelium, to compare the effects of iron supplied in physiologically relevant forms to either the apical or basolateral surfaces of the cells. Both sources of iron provoked increased cellular ferritin content, indicating iron uptake from both sides of the cells. Supply of basolateral transferrin-bound iron did not affect subsequent iron transport across the apical surface, but reduced iron transport across the basolateral membrane. In contrast, the apical iron supply led to subsequent reduction in iron transport across the apical cell membrane without altering iron export across the basolateral membrane. The apical and basolateral iron supplies also elicited distinct effects on the expression and subcellular distribution of iron transporters. These data suggest that, in addition to the effects of cellular iron status on the expression of iron transporter genes, different modes and direction of iron supply to enterocytes can elicit distinct functional effects on iron transport.

Electronic supplementary material

The online version of this article (doi:10.1007/s12263-015-0463-5) contains supplementary material, which is available to authorized users.     

配施磷细菌肥对复垦土壤细菌多样性及磷有效性的影响

为了解磷细菌肥对复垦土壤微生物群落结构变化特征和磷有效性的影响,本研究以连续配施磷细菌肥5年的定位试验为背景,利用16S rDNA基因序列测序方法对土壤细菌群落多样性进行分析,探讨土壤细菌群落与土壤Olsen-P、碱性磷酸酶的关系.试验共设对照、单施化肥、有机肥、有机肥化肥、化肥磷细菌肥、有机肥磷细菌肥和有机肥化肥磷细菌肥7个处理.结果表明: 复垦土壤中放线菌门和变形菌门菌群的相对丰度最大,分别为21.6％～32.2％和13.8％～28.9％.有机肥化肥磷细菌肥处理的OTU数和Chao1指数分别为809和26190,均属最高.磷细菌肥处理能提高土壤中放线菌门和变形菌门菌群的相对丰度,降低土壤中酸杆菌门、热袍菌门和硝化螺旋菌门菌群的相对丰度,对诺卡氏菌属、屈挠杆菌属有一定的促进作用.有机肥化肥磷细菌肥处理能够提高复垦土壤Olsen-P及碱性磷酸酶活性.复垦土壤变形菌门与Olsen-P、碱性磷酸酶的相关系数最高(0.900、0.955),在一定程度上可以作为土壤磷有效性的灵敏性指标.     

Conflicting effects of BMI and waist circumference on iron status

The association between obesity and iron status has a long history and is still receiving attention. However comparative analysis of the association between general obesity (BMI) and visceral obesity (waist circumference) with iron status has not been extensively researched. The aim of the present study is thus to determine if body mass index and waist circumference have the same correlation with iron status. One thousand one hundred and thirty people (225 men and 905 women) aged 30 years and above participated in this study. Anthropometric parameters, haemoglobin, iron and total iron binding capacity concentrations were measured using standard methods. Percentage transferrin saturation was calculated and ferritin concentrations were measured using an enzyme linked immunosorbent assay. Obese or overweight women had significantly lower iron and transferrin saturation concentration when compared to non-obese women. In contrast, women with high waist circumference had comparable plasma iron and transferrin saturation to women with normal waist circumference. Partial correlation analysis and linear regression analysis showed that BMI is negatively and significantly associated with plasma iron, transferrin saturation, Hb and ferritin concentration, whilst waist circumference is positively but insignificantly associated with plasma iron, transferrin saturation, Hb and ferritin concentration. Binary regression analysis showed that obese or overweight people are more likely to have iron deficiency, whilst those with raised waist circumference are more likely to have iron overload. Multivariate analysis showed that body mass index is negatively and significantly associated with low iron status, while waist circumference is positively and insignificantly associated with iron status. This is supported by a comparison of plasma iron, transferrin saturation and ferritin concentrations in participants with high body mass index and normal waist circumference and participants with normal body mass index and high waist circumference to those participants having normal body mass index and normal waist circumference. The present study suggests that in women body mass index is associated with low plasma iron, transferrin saturation and ferritin concentrations, while waist circumference is associated with high plasma iron, transferrin saturation and ferritin concentrations.     

Availability of iron,manganese, zinc and phosphorus in submerged sodic soil as affected by amendments during the growth period of rice crop

Summary Effect of amendments, gypsum (12.5 tonnes/ha), farmyard manure (30 tonnes/ha), rice husk (30 tonnes/ha) and also no amendment (control) on the availability of native Fe, Mn and P and applied Zn in a highly sodic soil during the growth period of rice crop under submerged conditions was studied in a field experiment. Soil samples were collected at 0, 30, 60 and 90 days of crop growth. Results showed that extractable Fe (1N NH₄OAC pH 3) and Mn (1N NH₄OAC pH 7) increased with submergence upto 60 days of crop growth but thereafter remained either constant or declined slightly. Application of farmyard manure and rice husk resulted in marked improvement of these elements over gypsum and control. Increases in extractable Mn (water soluble plus exchangeable) as a result of submergence and crop growth under different amendments were accompanied by corresponding decreases in easily reducible Mn content of the soil. Application of 40 kg zinc sulphate per hectare to rice crop could substantially raise the available Zn status (DTPA extractable) of the soil in gypsum and farmyard manure treated plots while the increase was only marginal in rice husk and control plots indicating greater fixation of applied Zn. Available P (0.5M NaHCO₃ pH 8.5) behaved quite differently and decreased in the following order with crop growth: gypsum>rice husk>farmyard manure>control.     

Influence of external zinc and phosphorus supply on Cd uptake by rice (Oryza sativa L.) seedlings with root surface iron plaque

Rice seedlings were grown in hydroponic culture to determine the effects of external Zn and P supply on plant uptake of Cd in the presence or absence of iron plaque on the root surfaces. Iron plaque was induced by supplying 50 mg l⁻¹ Fe²⁺ in the nutrient solution for 2 day. Then 43-day-old seedlings were exposed to 10 μmol l⁻¹ Cd together with 10 μmol l⁻¹ Zn or without Zn (Zn–Cd experiment), or to 10 μmol l⁻¹ Cd with 1.0 mmol l⁻¹ P or without P (P–Cd experiment) for another 2 day. The seedlings were then harvested and the concentrations of Fe, Zn, P and Cd in dithionite–citrate–bicarbonate (DCB) extracts and in roots and shoots were determined. The dry weights of roots and shoots of seedlings treated with 50 mg l⁻¹ Fe were significantly lower than when no Fe was supplied. Adsorption of Cd, Zn and P on the iron plaque increased when Fe was supplied but Cd concentrations in DCB extracts were unaffected by external Zn or P supply levels. Cd concentrations in shoots and roots were lower when Fe was supplied. Zn additions decreased Cd concentrations in roots but increased Cd concentrations in shoots, whereas P additions significantly increased shoot and root Cd concentrations and this effect diminished when Fe was supplied. The percentage of Cd in DCB extracts was significantly lower than in roots or shoots, accounting for up to 1.8–3.8% of the plant total Cd, while root and shoot Cd were within the ranges 57–76% and 21–40% respectively in the two experiments. Thus, the main barrier to Cd uptake seemed to be the root tissue and the contribution of iron plaque on root surfaces to plant Cd uptake was minor. The changes in plant Cd uptake were not due to Zn or P additions altering Cd adsorption on iron plaque, but more likely because Zn or P interfered with Cd uptake by the roots and translocation to the shoots.     

Pulses of soil phosphorus availability in a Mexican tropical dry forest: effects of seasonality and level of wetting

Intact cores from the upper soil profile and surface litter were collected at the peak of the dry season and during the rainy period in the tropical deciduous forest of the Chamela region, Jalisco, México, to (1) analyze upper soil phosphorus (P) movement and retention, (2) compare soil P dynamic pools (soluble, bicarbonate, and microbial) in dry and rainy seasons, and (3) determine the response of these P pools to wetting. Unperturbed litter-soil cores were treated in the laboratory with either 10 mm or 30 mm of simulated rain with carrier-free ³²P and compared to a control (no water addition) to determine the fate and retention of added P. ³¹P concentrations and pools in most litter and soil fractions were higher in the dry than in the rainy season. Soluble P was 0.306 g/m² and microbial P was 0.923 g/m² in the dry season (litter plus soil) versus 0.041 (soluble) and 0.526 (microbial) g P/m² in the rainy season. After water addition, rainy-season cores retained 99.9 and 94% of ³²P in the 10- and 30-mm treatments, respectively. Dry-season samples retained 98.9 and 80% of inputs in the same treatments. Retention after wetting occurred mostly in soil (bicarbonate and microbial fractions). Simulated rainfall on rainy-season soils increased P immobilization. On the other hand, simulated rainfall on dry-season soils released P through mineralization. The P release represents between 46 and 99% of the annual litterfall return. Our results suggest that both soluble and microbial P constitute important sources for initiation of plant growth at the onset of the rainy season in tropical dry forest. Received: 23 September 1997 / Accepted: 2 February 1998     

Environmental modulation of calcium and phosphorus metabolism in embryonic snapping turtles (Chelydra serpentina)

Summary Eggs of common snapping turtles (Chelydra serpentina) were incubated on wet (–150 kPa water potential) and dry (–950 kPa) substrates in a laboratory study assessing the effects of the hydric environment on patterns of mobilization of calcium and phosphorus by developing embryos. We found that embryos developing in wet environments withdrew nutrients from their yolk faster, grew more rapidly, and incubated longer than embryos exposed to dry environments. Turtles developing in both environments absorbed calcium from the yolk at similar rates and depleted the yolk of almost its entire reserve of calcium prior to hatching. Calcium withdrawn from the yolk was supplemented with calcium mobilized from the eggshell, but embryos in wet environments obtained substantially more calcium from the eggshell than did those in dry settings. Embryos obtained all of the phosphorus used in skeletogenesis from the yolk, but those incubating in wet environments mobilized phosphorus from this compartment more rapidly than did those in dry settings. Exposing embryonic snapping turtles to wet environments apparently allows them to make more efficient use of the transitory source of calcium in the eggshell than is possible in dry environments. However, the residual yolk in hatchlings from both wet and dry environments contains too little calcium to support the growth of hard and soft tissues in neonates at rates similar to those characterizing the growth phase of development in embryogenesis.     

Mechanisms for the increase in phosphorus uptake of waterlogged plants: soil phosphorus availability, root morphology and uptake kinetics

Waterlogging frequently reduces plant biomass allocation to roots. This response may result in a variety of alterations in mineral nutrition, which range from a proportional lowering of whole-plant nutrient concentration as a result of unchanged uptake per unit of root biomass, to a maintenance of nutrient concentration by means of an increase in uptake per unit of root biomass. The first objective of this paper was to test these two alternative hypothetical responses. In a pot experiment, we evaluated how plant P concentration of Paspalum dilatatum, (a waterlogging-tolerant grass from the Flooding Pampa, Argentina) was affected by waterlogging and P supply and how this related to changes in root-shoot ratio. Under both soil P levels waterlogging reduced root-shoot ratios, but did not reduce P concentration. Thus, uptake of P per unit of root biomass increased under waterlogging. Our second objective was to test three non-exclusive hypotheses about potential mechanisms for this increase in P uptake. We hypothesized that the greater P uptake per unit of root biomass was a consequence of: (1) an increase in soil P availability induced by waterlogging; (2) a change in root morphology, and/or (3) an increase in the intrinsic uptake capacity of each unit of root biomass. To test these hypotheses we evaluated (1) changes in P availability induced by waterlogging; (2) specific root length of waterlogged and control plants, and (3) P uptake kinetics in excised roots from waterlogged and control plants. The results supported the three hypotheses. Soil P avail-ability was higher during waterlogging periods, roots of waterlogged plants showed a morphology more favorable to nutrient uptake (finer roots) and these roots showed a higher physiological capacity to absorb P. The results suggest that both soil and plant mechanisms contributed to compensate, in terms of P nutrition, for the reduction in allocation to root growth. The rapid transformation of the P uptake system is likely an advantage for plants inhabiting frequently flooded environments with low P fertility, like the Flooding Pampa. This advantage would be one of the reasons for the increased relative abundance of P. dilatatum in the community after waterlogging periods. Received: 15 February 1997 / Accepted: 20 May 1997     

The composition and bioavailability of phosphorus transport through the Changjiang (Yangtze) River during the 1998 flood

Water and suspended sediment (SS) samples were collected from the Changjiang River at the Datong Hydrological Station (DHS), five times from May 1997 through January 1999 in order to evaluate transport, composition and bioavailability of phosphorus (P) during a 1998 flood. Transport of most of the phosphorus compositions was substantially higher during the 1998 flood than at other sampling dates. Phosphorus associated with suspended sediment (TPP) accounted for more than 85% of total phosphorus (TP) transport during periods of preflood and flood. The high transport of TPP during the flood was due to unusually high concentrations of TPP and sediment discharge. The potentially bioavailable phosphorus in SS (PBAP) accounted for about 10% of TPP. PBAP with dissolved inorganic phosphorus (DIP) consisted of 15–89% of TP. For all the sampling dates, the concentrations of potential bioavailable phosphorus (BAP) ranged from 0.035–0.08 mg L^–1, significantly higher than the limiting concentration for eutrophication. Therefore, the increasing temporal trends of TP concentration and high bioavailability of TP appear to support more frequent algal blooms in receiving East China Sea coastal waters in recent years. Hence, the underestimate of TPP transport by large rivers may also underestimate the biogeochemical cycling of other associated nutrients, such as nitrogen and carbon.     

Effect of iron limitation on protein composition and ultrastructure of Haemophilus ducreyi

Protein profiles of whole cells of Haemophilus ducreyi grown in the presence or absence of the iron chelator desferal, were compared by polyacrylamide gel electrophoresis. Each of four strains produced novel proteins in the range 43-160 kDa when cultured under conditions of reduced iron availability. At some sub-inhibitory concentrations, desferal produced enhanced growth, possibly due to it functioning as an exogenous siderophore. Organisms grown under conditions of reduced iron availability ultrastructurally showed also large periplasmic spaces between cytoplasm and outer membrane.     

Interaction of phosphorus and molybdenum and the availability of zinc,copper, manganese,molybdenum and phosphorus in waterlogged rice soil

Summary Laboratory incubation experiments were conducted with low-land rice soil to study the effect of applying three different levels of molybdenum (0, 2.5 and 5.0 ppm) and phosphorus (0, 100 and 200 ppm), in all possible combinations, on the changes in available Mo, P, Mn, Zn and Cu in soil. The results showed that application of Mo at both the levels increased the content of extractable Mo and P but decreased those of Cu and Mo in soil whereas application of Mo at higher level only increased the content of extractable Zn in soil. Application of P at both the levels decreased the content of extractable Mo, Mn and Cu but increased that of P whereas it showed an inconsistent effect on the extractable Zn content in soil. The P × Mo interaction effect was found to be beneficial for the content of P, Mo and Zn only. During the initial period of incubation all elements except Cu recorded an increase but with the progress of incubation period the content of all the elements except Mn gradually declined.     

Synergistic effect of Pseudomonas alkylphenolica PF9 and Sinorhizobium meliloti Rm41 on Moroccan alfalfa population grown under limited phosphorus availability

This study looked at the synergistic effect of Pseudomonas alkylphenolica PF9 and Sinorhizobium meliloti Rm41 on the Moroccan alfalfa population (Oued Lmaleh) grown under symbiotic nitrogen fixation and limited phosphorus (P) availability. The experiment was conducted in a growth chamber and after two weeks of sowing, the young seedlings were inoculated with Sinorhizobium meliloti Rm41 alone or combined with a suspension of Pseudomonas alkylphenolica PF9. Then, the seedlings were submitted to limited available P (insoluble P using Ca₃HPO₄) versus a soluble P form (KH₂PO₄) at a final concentration of 250 μmol P·plant⁻¹·week⁻¹. After two months of P stress, the experiment was evaluated through some agro-physiological and biochemical parameters. The results indicated that the inoculation of alfalfa plants with Sinorhizobium strain alone or combined with Pseudomonas strain significantly (p < 0.001) improved the plant growth, the physiological and the biochemical traits focused in comparison to the uninoculated and P-stressed plants. For most sets of parameters, the improvement was more obvious in plants co-inoculated with both strains than in those inoculated with Sinorhizobium meliloti Rm41 alone. In fact, under limited P-availability, the co-inoculation with two strains significantly (p < 0.01) enhanced the growth of alfalfa plants evaluated by fresh and dry biomasses, plant height and leaf area. The results indicated also that the enhancement noted in plant growth was positively correlated with the shoot and root P contents. Furthermore, the incensement in plant P contents in response to bacterial inoculation improved cell membrane stability, reflected by low malonyldialdehyde (MDA) and electrolyte leakage (EL) contents, and photosynthetic-related parameters such as chlorophyll contents, the maximum quantum yield of PS II (Fv/Fm) and stomatal conductance (g_s). Our findings suggest that Pseudomonas alkylphenolica PF9 can act synergistically with Sinorhizobium meliloti Rm41 in promoting alfalfa growth under low-P availability.     

Effects of time and dietary iron on tissue iron concentration as an estimate of relative bioavailability of supplemental iron sources for ruminants

Two experiments were conducted to investigate the effects of time and dietary Fe on tissue Fe concentrations following short-term, high level supplementation for use as a bioassay procedure for supplemental Fe sources for ruminants. In Experiment 1, 28 wethers were allotted randomly to four experimental diets which were fed for 15 or 30 days. The basal maize–soyabean meal–cottonseed hulls diet (193 mg kg⁻¹ Fe) was supplemented with 0, 400, 800 or1200 mg kg⁻¹ added Fe from reagent grade ferrous sulfate (FeSO₄·7H₂O). Iron concentrations in liver, kidney, and spleen increased (P<0.05) as dietary Fe increased; however, muscle, heart, and bone Fe concentrations were unaffected. A logarithmic transformation of liver or kidney Fe concentrations at 30 days regressed on added dietary Fe produced the best fits to a linear model. In Experiment 2, bioavailability of Fe from three feed grade ferrous carbonates known to differ (carbonates A, B, and C) was compared to that from reagent grade ferrous sulfate. The dietary treatments fed for 30 days included the above basal diet (90 mg kg⁻¹ Fe) supplemented with 0, 300, 600 or 900 mg kg⁻¹ added Fe from ferrous sulfate or 600 mg kg⁻¹ Fe from ferrous carbonates A, B, or C. Liver Fe concentrations from sheep fed ferrous sulfate were numerically greater than those of animals fed the carbonate sources or control diet. Kidney Fe concentrations from lambs fed ferrous sulfate at 600 mg kg⁻¹ Fe or carbonate-A were greater (P<0.05) than those fed carbonates B or C. Iron concentrations in spleen were lower (P<0.05) in lambs fed carbonate-B than for those fed 600 mg kg⁻¹ Fe as ferrous sulfate, but were similar to other carbonates. Overall average bioavailability estimates based on multiple regression slope ratios for the three tissues were ferrous sulfate 1.00, carbonate-A 0.55, carbonate-B 0.00, and carbonate-C 0.20. Estimates for carbonates A and C were similar to those based on hemoglobin concentrations reported previously for young swine supplemented at dietary concentrations near the requirement.     

Ca-independent effects of BAPTA and EGTA on single-channel Cl currents in brown adipocytes

The Cl⁻ channels of brown adipocytes electrophysiologically resemble outwardly rectifying Cl⁻ channels (ORCC). To study tentative Ca²⁺ regulation of these channels, we attempted to control Ca²⁺ levels at the cytoplasmic side of the inside-out membrane patches with Ca²⁺-chelating agents. However, we found that the commonly used Ca²⁺-chelators EGTA and BAPTA by themselves influenced the Cl⁻ channel currents, unrelated to their calcium chelating effects. Consequently, in this report we delineate effects of Ca²⁺-chelators (acting from the cytoplasmic side) on the single Cl⁻ channel currents in patch-clamp experiments. Using fixed (1-2 mM) concentrations of chelators, two types of Cl⁻ channels were identified, as discriminated by their reaction to the Ca²⁺-chelators and by their conductance: true-blockage channels (31 pS) and quasi-blockage channels (52 pS). In true-blockage channels, EGTA and BAPTA inhibited channel activity in a classical flickery type manner. In quasi-blockage channels, chelators significantly shortened the duration of individual openings, as in a flickering block, but the overall channel activity tended to increase. This dual effect of mean open time decrease accompanied by a tendency of open probability to increase we termed a quasi-blockage. Despite the complications due to the chelators as such, we could detect a moderate inhibitory effect of Ca²⁺. The anionic classical Cl⁻ channel blockers DIDS and SITS could mimic the true/quasi blockage of EGTA and BAPTA. It was concluded that at least in this experimental system, standard techniques for Ca²⁺ level control in themselves could fundamentally affect the behaviour of Cl⁻ channels.