Biodegradation of Tapis blended crude oil in marine sediment by a consortium of symbiotic bacteria

Biodegradation rate and the high molecular weight hydrocarbons are among the important concerns for bioremediation of crude oil. Inoculation of a non-oil-degrading bacterium as supplementary bacteria increased oil biodegradation from 57.1% to 63.0% after 10 days of incubation. Both the oil-degrading bacteria and the non-oil-degrading bacteria were isolated from Malaysian marine environment. Based on the 16S rDNA sequences, the oil-degrading bacteria was identified as Pseudomonas pseudoalcaligenes (99% similarity) while the non-oil-degrading bacterium was Erythrobacter citreus (99% similarity). E. citreus does not grow on crude oil enriched medium under present experimental condition but it withstands 5000 mg kg^?1 Tapis blended crude oil in sediment. Under optimal condition, the oil-degrading bacterium; P. pseudoalcaligenes, alone utilized 583.3 ± 3.8 mg kg^?1 (57.1%) at the rate of 3.97 × 10^?10 mg kg^?1 cell^?1 day^?1 Tapis blended crude oil from 1000 mg kg^?1 oil-contaminated sediment. Inoculation of E. citreus as the supplementary bacteria to P. pseudoalcaligenes enhanced biodegradation. The bacterial consortium degraded 675.8 ± 18.5 mg kg^?1 (63.0%) Tapis blended crude oil from the 1000 mg kg^?1 oil-contaminated sediment. Biodegradation rate of the bacterial consortium increased significantly to 4.59 × 10^?10 mg kg^?1 cell^?1 day^?1 (p = 0.02). Improvement of the oil degradation by the bacterial consortium was due to the synergetic reaction among the bacterial inoculants. There are two implications: (1) E. citreus may have a role in removing self-growth-inhibiting compounds of P. pseudoalcaligens. (2) P. pseudoalcaligenes degraded Tapis blended crude oil while E. citreus competes for the partially degraded hydrocarbons by P. pseudoalcaligenes. P. pseudoalcaligenes forced to breakdown more hydrocarbons to sustain its metabolic requirement. The bacterial consortium degraded 78.7% of (C₁₂–C₃₄) total aliphatic hydrocarbons (TAHs) and 74.1% of the 16 USEPA prioritized polycyclic aromatic hydrocarbons.     

Effect of the dietary probiotic Clostridium butyricum on growth,intestine antioxidant capacity and resistance to high temperature stress in kuruma shrimp Marsupenaeus japonicus

A 56-day feeding trial followed by an acute high temperature stress test were performed to evaluate the effect of dietary probiotic Clostridium butyricum (CB) on growth performance and intestine antioxidant capacity of kuruma shrimp Marsupenaeus japonicus. Shrimp were randomly allocated in 9 tanks (30 shrimp per tank) and triplicate tanks were fed with diets containing different levels of C. butyricum (1×10⁹ cfu/g): 0 mg g⁻¹ feed (Control), 100 mg g⁻¹ feed (CB-100), 200 mg g⁻¹ feed (CB-200) as treatment groups. The results indicated that dietary supplementation of C. butyricum increased the growth performance and decreased the feed conversion rate (FCR) of shrimp in the CB-100 group. HE stain showed that C. butyricum increased the intestine epithelium height of M. japonicus. C. butyricum supplemented in diets decreased·O₂^- generation capacity and malondialdehyde (MDA) content, and increased total antioxidant capacity (T-AOC), catalase (CAT) and peroxidase (POD) activity and the expression level of heat shock protein 70 (hsp70) and metallothionein (mt) gene in intestine of shrimp cultured under normal condition for 56 d, while no significant changes in glutathione peroxidase (GPx) activity and ferritin gene expression level. After shrimp exposed to high temperature stress 48 h, the lower level of·O₂^- generation capacity and MDA content, and the higher level survival, activities of T-AOC, CAT, GPx and POD, as well as hsp70, ferritin and mt gene expression level were found in intestine of two C. butyricum groups. These results revealed that C. butyricum could improve the growth performance, increase intestine antioxidant capacity of M. japonicus against high temperature stress, and could be a potential feed additive in shrimp aquaculture.     

Antioxidative,antimicrobial and cytotoxic effects of the phenolics of Leea indica leaf extract

This study investigated the phytochemical, antioxidative, antimicrobial and cytotoxic effects of Leea indica leaf ethanol extract. Phytochemical values namely total phenolic and flavonoid contents, total antioxidant capacity, DPPH radical scavenging effect, FeCl₃ reducing power, DMSO superoxide scavenging effect and Iron chelating effects were studied by established methods. Antibacterial, antifungal and cytotoxic effects were screened by disk diffusion technique, food poison technique and brine shrimp bioassay, respectively. Results showed the total phenolic content 24.00 ± 0.81 g GAE/100 g, total flavonoid content 194.68 ± 2.43 g quercetin/100 g and total antioxidant capacity 106.61 ± 1.84 g AA/100 g dry extract. Significant (P < 0.05) IC₅₀ values compared to respective standards were recorded in DPPH radical scavenging (139.83 ± 1.40 μg/ml), FeCl₃ reduction (16.48 ± 0.64 μg/ml), DMSO superoxide scavenging (676.08 ± 5.80 μg/ml) and Iron chelating (519.33 ± 16.96 μg/ml) methods. In antibacterial screening, the extract showed significant (P < 0.05) zone of inhibitions compared to positive controls Ampicillin and Tetracycline against Gram positive Bacillus subtilis, Bacillus cereus, Bacillus megaterium, and Staphylococcus aureus and Gram negative Salmonella typhi, Salmonella paratyphi, Pseudomonas aeroginosa, Shigella dysenteriae, Vibrio cholerae, and Escherichia coli. Significant minimum inhibitory concentrations compared to tetracycline were obtained against the above organisms. In antifungal assay, the extract inhibited the growth of Aspergillus flavus, Candida albicans and Fusarium equisetii by 38.09 ± 0.59, 22.58 ± 2.22, and 22.58 ± 2.22%, respectively. The extract showed a significant LC₅₀ value compared to vincristine sulfate in cytotoxic assay. The results evidenced the potential antioxidative, antimicrobial and cytotoxic capacities of Leea inidica leaf extract to be processed for pharmaceutical use.     

Selenium uptake,speciation and stressed response of Nicotiana tabacum L.

The research on the function and mechanism of selenium (Se) is of great significance for the development of Se-enriched agricultural products. In this paper, uptake, speciation distribution, the effects on the flue-cured tobacco growth and antioxidant system of Se at different levels (0–22.2 mg Se kg⁻¹) were studied through a pot experiment, aiming to clarify flue-cured tobacco's response to Se stress and the relationship between Se speciation and antioxidant system. The results showed that the leaf area and number, the biomass and the chlorophyll content reached the maximum at 4.4 mg kg⁻¹ of Se treatment. Selenium at low levels (≤4.4 mg kg⁻¹) stimulated the growth of flue-cured tobacco by elevating the capability of antioxidant stress and reducing the malondialdehyde (MDA) content to 0.6–0.8 times of that of the control. However, high Se levels (≥11.1 mg kg⁻¹) depressed the capability of antioxidant stress and raised the MDA content to 1.5-fold of that of the control, and meanwhile the biomass of the aboveground parts and underground parts declined notably. The Se content in different parts of flue-cured tobacco significantly increased with the growth of Se levels. The range of Se content in roots, leaves and stems at 2.2–22.2 mg kg⁻¹ of Se treatment were 16.7–58.6 mg kg⁻¹, 2.6–37.3 mg kg⁻¹ and 2.2–10.3 mg kg⁻¹, respectively. According to the detection of different Se speciation, only selenocysteine (SeCys) was detectable in leaves at 2.2 mg kg⁻¹ Se treatment; SeCys, selenite [Se(IV)]and selenate [Se(VI)] were detected in flue-cured tobacco leaves at Se treatment (≥4.4 mg kg⁻¹), which accounted for 4.6–10%, 9–18.7% and 71–86% respectively; SeCys, selenomethionine (SeMet) and Se(IV) were detected in roots, and organic selenium(66–84%) was the main Se species at Se ≤ 11.1 mg kg⁻¹ treatment; four Se species [SeCys, SeMet, Se(IV) and Se(VI)] were detected in flue-cured tobacco roots, and the main Se species was inorganic Se (60%) at 22.2 mg kg⁻¹ Se treatment. That was to say, the percentage of organic Se species (SeCys and SeMet in flue-cured tobacco leaves and root) declined, whereas the ratio of inorganic Se species [Se(IV) and Se(VI)] increased with the growth of Se levels. The correlation analysis showed that the superoxide dismutase (SOD) activity as well as the glutathione (GSH) and MDA contents were positively correlated with the Se(IV) and Se(VI) contents at P < 0.01 and excessive inorganic Se might destruct the reactive oxygen species (ROS) balance and enhance the MDA content, thus causing damage to the plant growth. In a word, the present study suggested that the ratio of inorganic Se [Se(IV) and Se(VI)] was closely related with the growth and the antioxidant capacity of flue-cured tobacco and the excessive application of Se led to the higher proportion of inorganic Se and poorer antioxidant capacity, which ultimately inhibited the growth of flue-cured tobacco.     

Response of native grasses and Cicer arietinum to soil polluted with mining wastes: Implications for the management of land adjacent to mine sites

Mine tailings are an environmental problem in Southern Spain because wind and water erosion of bare surfaces results in the dispersal of toxic metals over nearby urban or agricultural areas. Revegetation with tolerant native species may reduce this risk. We grew two grasses, Lygeum spartum and Piptatherum miliaceum, and the crop species Cicer arietinum (chickpea) under controlled conditions in pots containing a mine tailings mixed into non-polluted soil to give treatments of 0%, 25%, 50%, 75% and 100% mine tailings. We tested a neutral (pH 7.4) mine tailings which contained high concentrations of Cd, Cu, Pb and Zn. Water-extractable metal concentrations increased in proportion to the amount of tailings added. The biomass of the two grasses decreased in proportion to the rate of neutral mine-tailing addition, while the biomass of C. arietinum only decreased in relation to the control treatment. Neutron radiography revealed that root development of C. arietinum was perturbed in soil amended with the neutral tailings compared to those of the control treatment, despite a lack of toxicity symptoms in the shoots. In all treatments and for all metals, the plants accumulated higher concentrations in the roots than in shoots. The highest concentrations occurred in the roots of P. miliaceum (2500 mg kg^?1 Pb, 146 mg kg^?1 Cd, 185 mg kg^?1 Cu, 2700 mg kg^?1 Zn). C. arietinum seeds had normal concentrations of Zn (70–90 mg kg^?1) and Cu (6–9 mg kg^?1). However, the Cd concentration in this species was ～1 mg kg^?1 in the seeds and 14.5 mg kg^?1 in shoots. Consumption of these plant species by cattle and wild fauna may present a risk of toxic metals entering the food chain.     

Evaluation of phosphorus distribution and bioavailability in sediments of a subtropical wetland reserve in southeast China

The phosphorus (P) fractions and bioavailable P in the sediments from the Quanzhou Bay Estuarine Wetland Nature Reserve were investigated using chemical extraction methods for the first time to study the distribution and bioavailability of P in the reserve sediments. A hypothesis was presented suggesting that the bioavailable P in the sediments could be evaluated using the P fractions. The total phosphorus (TP), inorganic phosphorus (IP), organic phosphorus (OP), non-apatite phosphorus (NAIP), and apatite phosphorus (AP) contents in the sediments were in the ranges of 303.87–761.59 mg kg⁻¹, 201.22–577.66 mg kg⁻¹, 75.83–179.16 mg kg⁻¹, 28.86–277.90 mg kg⁻¹, and 127.36–289.94 mg kg⁻¹, respectively. The water soluble phosphorus (WSP), readily desorbable phosphorus (RDP), algal available phosphorus (AAP), and NaHCO₃ extractable phosphorus (Olsen-P) contents in the sediments were in the ranges of 0.58–357.17 mg kg⁻¹, 80.77–586.75 mg kg⁻¹, 1.09–24.12 mg kg⁻¹, and 54.96–676.82 mg kg⁻¹, respectively. The correlation analysis results showed that the NAIP was the major component of the bioavailable P and that the impact of the AP on the bioavailable phosphorus may be minimal. Due to the low TP content in the sediments of the Quanzhou Bay Estuarine Wetland Nature Reserve, the potential pollution risks of P in the sediments may not be very high. The results also show that the bioavailable P concentrations in the sediments of the Quanzhou Bay Estuarine Wetland Nature Reserve could not be evaluated by measuring the P fractions and that the hypothesis was untenable.     

Recycling of agro-industrial sludge through vermitechnology

This work illustrates the feasibility of vermitechnology to stabilize sludge from an agro-industry. To achieve the goal, industrial sludge (IS) was mixed with three different bulky agents, i.e. cow dung (CD), biogas plant slurry (BGS) and wheat straw (WS), in different ratios to produce nine different feed mixtures for earthworm Eisenia fetida. Vermicomposting bedding material was analyzed for its different physic-chemical parameters after 15 weeks of experimentations. In all waste mixtures, a decrease in pH, organic C and C:N ratio, but increase in total N, available P, exchangeable K, exchangeable Ca and trace elements (Mg, Fe and Zn) was recorded. IS (40%) + CD (60%) and IS (40%) + BGS (60%) vermibeds showed the highest mineralization rate and earthworm growth patterns during vermicomposting process. Vermicompost contains (dry weight basis) a considerable range of plant available forms of P (17.5–28.9 g kg^?1), K (13.8–21.4 g kg^?1), Ca (41.1–63.4 g kg^?1), Mg (262.4–348.3 mg kg^?1), Fe (559.8–513.0 mg kg^?1) and Zn (363.1–253.6 mg kg^?1). Earthworm growth parameters, i.e. biomass gain, total cocoon production, individual growth rate (mg wt. worm^?1 day^?1), natality rate, total fecundity were optimum in bedding containing 20–40% industrial sludge. C:N ratio of worm-processed material was within the agronomic acceptable or favorable limit (<15–20). The results clearly suggested that vermitechnology can be a potential technology to convert industrial sludges into vermifertilizer for sustainable land restoration practices.     

Arsenic reduced scale-insect infestation on arsenic hyperaccumulator Pteris vittata L.

Arsenic hyperaccumulation by Pteris vittata L. (Chinese brake fern) may serve as a defense mechanism against herbivore attack. This study examined the effects of arsenic exposure (0, 5, 15 and 30 mg kg^?1) on scale insect (Saissetia neglecta) infestation of P. vittata. Scale insects were counted as a percentage fallen from the plant to the total number of insects after 1 week of As-treatment. The arsenic concentrations in the fronds ranged from 5.40 to 812 mg kg^?1. Greater arsenic concentrations resulted in higher percentage of fallen-scale insects (17.2–55.0%). Lower arsenic concentrations (≤5 mg kg^?1) showed significantly lower effect on the population compared to 15–30 mg kg^?1 (p < 0.05). Arsenic content in the fallen-scale insects was as high as 194 mg kg^?1, which indicated that arsenic has been ingested by the scale insects via plant sap. This study is consistent with the hypothesis that arsenic may help P. vittata defend against herbivore's attack.     

Lead,zinc, cadmium hyperaccumulation and growth stimulation in Arabis paniculata Franch

Metal hyperaccumulation is of great interest in recent years because of its potential application for phytoremediation of heavy metal contaminated soils. In this study, a field survey and a hydroponic experiment were conducted to study the accumulation characteristics of lead (Pb), zinc (Zn) and cadmium (Cd) in Arabis paniculata Franch., which was found in Yunnan Province, China. The field survey showed that the wild population of A. paniculata was hyper-tolerant to extremely high concentrations of Pb, Zn and Cd, and could accumulate in shoots an average level of 2300 mg kg^?1 dry weight (DW) Pb, 20,800 mg kg^?1 Zn and 434 mg kg^?1 Cd, with their translocation factors (TFs) all above one. Under the hydroponic culture, stimulatory effects of Pb, Zn and Cd on shoot dry biomass were noted from 24 to 193 μM Pb, 9 to 178 μM Cd and all Zn supply levels in nutrient solution, while the effects were not obvious in the roots. Chlorophyll concentrations in Pb, Zn and Cd treatments showed an inverted U-shaped pattern, consistent with the change of plant biomass. Pb, Zn and Cd concentrations in the shoots and roots increased sharply with increasing Pb, Zn and Cd supply levels. They reached > 1000 mg kg^?1 Pb, 10,000 mg kg^?1 Zn and 100 mg kg^?1 Cd DW in the 24 μM Pb, 1223 μM Zn and 9 μM Cd treatment, respectively, in which the plants grew healthy and did not show any symptoms of phytotoxicity. The TFs of Zn were basically higher than one and the amount of Zn taken by shoots ranged from 78.7 to 90.4% of the total Zn. However, the TFs of Pb and Cd were well below one, and 55.0–67.5% of total Pb and 57.8–83.5% of total Cd was accumulated in the shoots. These results indicate that A. paniculata has a strong ability to tolerate and hyperaccumulate Pb, Zn and Cd. Meanwhile, suitable levels of Pb, Zn and Cd could stimulate the biomass production and chlorophyll concentrations of A. paniculata. Thus, it provides a new plant material for understanding the mechanisms of stimulatory effect and co-hyperaccumulation of multiple heavy metals.     

Recovery of protein,chitin, carotenoids and glycosaminoglycans from Pacific white shrimp (Litopenaeus vannamei) processing waste

Shrimp head waste is a major byproduct of crustacean processing in North-eastern Brazil and represents an interesting source of bioactive molecules. Additionally, its use increases the sustainability of processing fishery products. The present study reports a process developed for recovering bioactive molecules from shrimp heads through autolysis. A protein hydrolysate (120 ± 0.4 g) formed by a 9% (w/v) solution was recovered and lyophilized from 1 kg of shrimp heads. Approximately 195 ± 0.5 mg of carotenoids was recovered as an ethanolic extract. The recovery of chitin and chitosan were 25 ± 2 g kg^?1 and 17 ± 4 g kg^?1 wet processing waste, respectively. Chitosans were characterized by ¹³C NMR, and FT-IR analysis and exhibited a variable degree of deacetylation (60–80%). Sulfated glycosaminoglycans that exhibited electrophoretic migration similar to mammalian standards were also recovered (79 ± 2 mg kg^?1 wet processing waste), and their degradation products suggested the presence of C6-sulfated heparan sulfate. These data point to the feasibility of an integrated process for isolating highly bioactive molecules, such as sulfated- and amino-polysaccharides, with a broad spectrum of applications from shrimp processing waste.     

Trace amounts of nickel in belowground rhizomes of Vaccinium myrtillus L. decrease anthocyanin concentrations in aerial shoots without water stress

Nickel (Ni) may impair plant water balance through detrimental effects on the belowground level. Bilberry (Vaccinium myrtillus L.) plants were grown in a mesic heath forest-type soil and subjected to Ni sulphate (NiSO₄·6H₂O) concentrations of 0, 10, 50, 100 and 500 mg m⁻² during an entire growing season in northern Finland (65°N). Biomass of belowground rhizomes, and tissue water content (TWC) and anthocyanin concentrations of aerial shoots were determined from mature plants in order to study rhizospheric Ni stress, and its possible long-distance effects on aerial shoots. As the major proportion of biomass of bilberry is invested in belowground parts, it was hypothesised that Ni-induced rhizospheric disturbance causes water stress in aerial shoots and increases their anthocyanin concentrations for osmotic regulation. Uptake of Ni from the soil to the rhizome and aerial shoots was measured with X-ray fluorescence spectrometry. Ni concentrations in the soil and rhizome exhibited a dose–response relationship, but the concentrations in the rhizome were about 10-fold lower (<3 mg Ni kg⁻¹) than those in the soil (<30 mg Ni kg⁻¹). Translocation of Ni from the rhizome to aerial shoots did not occur, as Ni concentrations in shoots remained at 1 mg Ni kg⁻¹. Although Ni concentrations in the rhizome were below the threshold values of Ni toxicity (i.e. 10–50 mg Ni kg⁻¹), Ni decreased the rhizome biomass. Anthocyanins decreased in aerial shoots along with the Ni accumulation in the rhizome, while TWC was unaffected. The result suggests that anthocyanins are not involved in osmotic regulation under Ni stress, since anthocyanins in aerial shoots responded to the Ni concentrations in the rhizome despite the lack of water stress.     

Reducing nutrient loss from farms through silvopastoral practices in coarse-textured soils of Florida,USA

Trees integrated into the range- and pasturelands of Florida could remove nutrients from deeper soil profiles that would otherwise be transported to water bodies and cause pollution. Soil nitrogen (N) and phosphorus (P) concentrations were monitored in three pastures: a treeless pasture of bahiagrass (Paspalum notatum); a pasture of bahiagrass under 20-year-old slash pine (Pinus elliotti) trees (silvopasture); and a pasture of native vegetation under pine trees (native silvopasture). Soil analysis from 10 profiles within each pasture showed that P concentrations were higher in treeless pasture (mean: 9.11 mg kg⁻¹ in the surface to 0.23 mg kg⁻¹ at 1.0 m depth) compared to silvopastures (mean: 2.51 and 0.087 mg kg⁻¹, respectively), and ammonium–N and nitrate–N concentrations were higher in the surface horizon of treeless pasture. The more extensive rooting zones of the combined stand of tree + forage may have caused higher nutrient uptake from silvopastures than treeless system. Further, compared to treeless system, soils under silvopasture showed higher P storage capacity. The results suggest that, compared to treeless pasture, silvopastoral association enhances nutrient retention in the system and thus reduces chances for nutrient transport to surface water. The study reflects the scope for applying ecological-engineering and ecosystem-restoration principles to silvopastoral-system design.     

Free radical scavenging activity from different extracts of leaves of Bauhinia vahlii Wight & Arn.

The objectives of this study were to determine phenolic content and antioxidant activities of chloroform, acetone, methanol and hot water extracts of Bauhinia vahlii leaves. The hot water extract afforded the highest yield (6.3%) while the lowest yield was obtained from the chloroform extract (2.1%). The methanol extract contains higher levels of total phenolics (48.7 ± 0.7 g GAE/100 g extract), tannins (21.7 ± 0.7 g GAE/100 g extract) and flavonoids (10.3 ± 0.2 RE/100 g extract). The extracts were subjected to assess their antioxidant potential using various in vitro systems such as DPPH, ABTS⁺, FRAP, OH, β-carotene linoleic acid bleaching system, phosphomolybdenum reduction and Fe²⁺ chelation. It is concluded that the methanolic extract of B. vahlii leaves have strong antioxidant potential. Further study is necessary for isolation and characterization of the active antioxidants, which may serve as a potential source of natural antioxidants.     

In vivo antimicrobial evaluation of an alanine-rich peptide derived from Pleuronectes americanus

In several organisms, the first barrier against microbial infections consists of antimicrobial peptides (AMPs) which are molecules that act as components of the innate immune system. Recent studies have demonstrated that AMPs can perform various functions in different tissues or physiological conditions. In this view, this study was carried out in order to evaluate the multifunctional activity in vivo of an alanine-rich peptide, known as Pa-MAP, derived from the polar fish Pleuronectes americanus. Pa-MAP was evaluated in intraperitoneally infected mice with a sub-lethal concentration of Escherichia coli at standard concentrations of 1 and 5 mg kg⁻¹. At both concentrations, Pa-MAPs exhibited an ability to prevent E. coli infection and increase mice survival, similar to the result observed in mice treated with ampicillin at 2 mg kg⁻¹. In addition, mice were monitored for weight loss. The results showed that mice treated with Pa-MAPs at 1 mg kg⁻¹ gained 0.8% of body weight during the 72 h of experiment. The same was observed with Pa-MAP at 5 mg kg⁻¹, which had a gain of 0.5% in body weight during the treatment. Mice treated with ampicillin at 2 mg kg⁻¹ show a significant weight loss of 5.6% of body weight. The untreated group exhibited a 5.5% loss of body weight. The immunomodulatory effects were also evaluated by the quantification of IL-10, IL-12, TNF-α, IFN-γ and nitric oxide cytokines in serum, but no immunomodulatory activity was observed. Data presented here suggest that Pa-MAP should be used as a novel antibiotic against infection control.     

Protective effect of Piper betle leaf extract against cadmium-induced oxidative stress and hepatic dysfunction in rats

The present study was undertaken to examine the attenuative effect of Piper betle leaf extract (PBE) against cadmium (Cd) induced oxidative hepatic dysfunction in the liver of rats. Pre-oral supplementation of PBE (200 mg/kg BW) treated rats showed the protective efficacy against Cd induced hepatic oxidative stress. Oral administration of Cd (5 mg/kg BW) for four weeks to rats significantly (P > 0.05) elevated the level of serum hepatic markers such as serum aspartate transaminase (AST), serum alanine transaminase (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), gamma-glutamyl transpeptidase (GGT), bilirubin (TBRNs), oxidative stress markers viz., thiobarbituric acid reactive substances (TBARS), lipid hydroperoxides (LOOH), protein carbonyls (PC) and conjugated dienes (CD) and significantly (P > 0.05) reduced the enzymatic antioxidants viz., superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione S-transferase (GST), glutathione reductase (GR) and glucose-6-phosphate dehydrogenase (G6PD) and non-enzymatic antioxidants Viz., reduced glutathione (GSH), total sulfhydryls (TSH), vitamin C and vitamin E in the liver. Pre-oral supplementation of PBE (200 mg/kg BW) in Cd intoxicated rats, the altered biochemical indices and pathological changes were recovered significantly (P > 0.05) which showed ameliorative effect of PBE against Cd induced hepatic oxidative stress. From the above findings, we suggested that the pre-administration of P. betle leaf extract exhibited remarkable protective effects against cadmium-induced oxidative hepatic injury in rats.     

Mathematical analysis of trickling filter response to pentachlorophenol shock load

The response of a laboratory trickling filter to a step increase in pentachlorophenol (PCP) feed concentration was analyzed using continuous stirred tank (CSTR) and plug flow reactor (PFR) models. The CSTR model provided a slightly better fit to experimental data than the plug flow model when specific growth rate, μ, and PCP-degrading biomass concentration before the shock load, X₀, were variable parameters but was clearly superior when the mean residence time, τ, was added as a third parameter. The three-parameter CSTR model accurately represented six of seven concentration response curves corresponding to step increases in PCP feed concentration of 12–165 mg l⁻¹ and 20–150 mg l⁻¹. The continuing improvement in system response to repetitive 20–150 mg l⁻¹ shock loads was reflected by a monotonic increase in the optimal estimates of initial rate of biomass production.     

Iodine concentration of milk in a dose–response study with dairy cows and implications for consumer iodine intake

Most feed is poor in iodine and iodine supplementation of cow's diets must guarantee milk iodine concentrations for humans that contribute to prevention of the deficiency and minimize the risk of exceeding an upper limit of iodine intake. Five Holstein cows were fed four iodine doses (via Ca(ΙO₃)₂·6H₂O). In four sequential 14-d periods, doses of 0.2 (basal diet), 1.3, 5.1, and 10.1 mg iodine kg^?1 diet dry matter (DM) were administered. Samples of milk were collected during each period; blood was also sampled from each cow for each iodine dosage. In an 18-d depletion period, a non-supplemented diet was provided. Iodine was determined by inductively coupled plasma-mass spectrometry. The iodine content of milk and serum reflected the iodine dosages in feed significantly. The levels for the four doses tested in milk were 101±32, 343±109, 1215±222, and 2762±852 μg iodine kg^?1. The total amount of iodine in milk per day was 30–40% of ingested supplemental iodine. Omitting additional iodine resulted in a short-term reduction of serum and milk iodine following an exponential decay function. The iodine supplementation of 0.5–1.5 mg kg^?1 diet DM represents the requirement of the cow, resulting in 100–300 μg iodine L^?1 milk, which optimally contributes to human supply. The maximum dietary levels of former and present EU legislations (10 and 5 mg iodine kg^?1 cow feed) increase the risk of iodine excess in humans.     

Antinociceptive action of NOP and opioid receptor agonists in the mouse orofacial formalin test

Nociceptin/orphanin FQ (N/OFQ) modulates several biological functions, including pain transmission via selective activation of a specific receptor named NOP. The aim of this study was the investigation of the antinociceptive properties of NOP agonists and their interaction with opioids in the trigeminal territory. The orofacial formalin (OFF) test in mice was used to investigate the antinociceptive potential associated to the activation of NOP and opioid receptors. Mice subjected to OFF test displayed the typical biphasic nociceptive response and sensitivity to opioid and NSAID drugs. Mice knockout for the NOP gene displayed a robust pronociceptive phenotype. The NOP selective agonist Ro 65-6570 (0.1–1 mg kg⁻¹) and morphine (0.1–10 mg kg⁻¹) elicited dose dependent antinociceptive effects in the OFF with the alkaloid showing larger effects; the isobologram analysis of their actions demonstrated an additive type of interaction. The mixed NOP/opioid receptor agonist cebranopadol elicited potent (0.01–0.1 mg kg⁻¹) and robust antinociceptive effects. In the investigated dose range, all drugs did not modify the motor performance of the mice in the rotarod test. Collectively the results of this study demonstrated that selective NOP agonists and particularly mixed NOP/opioid agonists are worthy of development as innovative drugs to treat painful conditions of the trigeminal territory.     

Tolerance,accumulation and distribution of zinc and cadmium in hyperaccumulator Potentilla griffithii

In this study, zinc (Zn) and cadmium (Cd) tolerance, accumulation and distribution was conducted in Potentilla griffithii H., which has been identified as a new Zn hyperaccumulator found in China. Plants were grown hydroponically with different levels of Zn²⁺ (20, 40, 80 and 160 mg L^?1) and Cd²⁺ (5, 10, 20 and 40 mg L^?1) for 60 days. All plants grew healthy and attained more biomass than the control, except 40 mg L^?1 Cd treatment. Zn or Cd concentration in plants increased steadily with the increasing addition of Zn or Cd in solution. The maximum metal concentrations in roots, petioles and leaves were 14,060, 19,600 and 11,400 mg kg^?1 Zn dry weight (DW) at 160 mg L^?1 Zn treatment, and 9098, 3077 and 852 mg kg^?1 Cd DW at 40 mg L^?1 Cd treatment, respectively. These results suggest that P. griffithii has a high ability to tolerate and accumulate Cd and Zn, and it can be considered not only as Zn but also as a potential cadmium hyperaccumulator. Light microscope (LM) with histochemical method, scanning electron microscope combined with energy dispersive spectrometry (SEM-EDS) and transmission electron microscope (TEM) were used to determine the distribution of Zn and Cd in P. griffithii at tissue and cellular levels. In roots, SEM-EDS confirmed that the highest Zn concentration was found in xylem parenchyma cells and epidermal cells, while for Cd, a gradient was observed with the highest Cd concentration in rhizodermal and cortex cells, followed by central cylinder. LM results showed that Zn and Cd distributed mainly along the walls of epidermis, cortex, endodermis and some xylem parenchyma. In leaves, Zn and Cd shared the similar distribution pattern, and both were mostly accumulated in epidermis and bundle sheath. However, in leaves of 40 mg L^?1 Cd treatment, which caused the phytotoxicity, Cd was also found in the mesophyll cells. The major storage site for Zn and Cd in leaves of P. griffithii was vacuoles, to a lesser extent cell wall or cytosol. The present study demonstrates that the predominant sequestration of Zn and Cd in cell walls of roots and in vacuoles of epidermis and bundle sheath of leaves may play a major role in strong tolerance and hyperaccumulation of Zn and Cd in P. griffithii.     

GLP-1(32–36)amide,a novel pentapeptide cleavage product of GLP-1, modulates whole body glucose metabolism in dogs

We have previously demonstrated in human subjects who under euglycemic clamp conditions GLP-1(9–36)amide infusions inhibit endogenous glucose production without substantial insulinotropic effects. An earlier report indicates that GLP-1(9–36)amide is cleaved to a nonapeptide, GLP-1(28–36)amide and a pentapeptide GLP-1(32–36)amide (LVKGR amide). Here we study the effects of the pentapeptide on whole body glucose disposal during hyperglycemic clamp studies. Five dogs underwent indwelling catheterizations. Following recovery, the dogs underwent a 180 min hyperglycemic clamp (basal glucose +98 mg/dl) in a cross-over design. Saline or pentapeptide (30 pmol kg⁻¹ min⁻¹) was infused during the last 120 min after commencement of the hyperglycemic clamp in a primed continuous manner. During the last 30 min of the pentapeptide infusion, glucose utilization (M) significantly increased to 21.4 ± 2.9 mg kg⁻¹ min⁻¹compared to M of 14.3 ± 1.1 mg kg⁻¹ min⁻¹ during the saline infusion (P = 0.026, paired t-test; P = 0.062, Mann–Whitney U test). During this interval, no significant differences in insulin (26.6 ± 3.2 vs. 23.7 ± 2.5 μU/ml, P = NS) or glucagon secretion (34.0 ± 2.1 vs. 31.7 ± 1.8 pg/ml, P = NS) were observed. These findings demonstrate that under hyperglycemic clamp studies the pentapeptide modulates glucose metabolism by a stimulation of whole-body glucose disposal. Further, the findings suggest that the metabolic benefits previously observed during GLP-1(9–36)amide infusions in humans might be due, at least in part, to the metabolic effects of the pentapeptide that is cleaved from the pro-peptide, GLP-1(9–36)amide in the circulation.