Toxicity of Crude Extracellular Products of <Emphasis Type="Italic">Aeromonas hydrophila</Emphasis> on Rohu, <Emphasis Type="Italic">Labeo rohita</Emphasis> (Ham.)

In the present study the haemolytic and proteolytic activity of extracellular products (ECP) secreted from Aeromonas hydrophila (CAHH14 strain) were studied with respect to temperature and different time of incubation as well as its lethal toxicity on rohu, Labeo rohita. The strain was isolated from Catla catla (showing abdominal dropsy symptom) collected from the pond of Central Institute of Freshwater Aquaculture (CIFA), Bhubaneswar, India and was characterized on the basis of biochemical tests. The highest production of haemolysin was achieved when the bacteria was grown at 35°C for 30 h. The proteolytic activity was found to be highest when the bacterium was grown at 30°C for 36 h. The haemolytic and proteolytic toxin produced by Aeromonas hydrophila was found to be lethal to rohu (LD₅₀ 1.7 × 10⁴ cfu/ml). The lethality of ECP was decreased by heating and completely inactivated by boiling at 100°C for 10 min. This indicates that protease activity and haemolytic activity of A. hydrophila ECP was temperature dependant.     

An eukaryotic translation initiation factor,AteIF5A‐2, affects cadmium accumulation and sensitivity in Arabidopsis

Cadmium (Cd) is one of the most toxic elements and can be accumulated in plants easily; meanwhile, eIF5A is a highly conserved protein in all eukaryotic organisms. The present work tried to investigate whether eIF5A is involved in Cd accumulation and sensitivity in Arabidopsis (Arabidopsis thaliana L.) by comparing the wild‐type Columbia‐0 (Col‐0) with a knockdown mutant of AteIF5A‐2, fbr12‐3 under Cd stress conditions. The results showed that the mutant fbr12‐3 accumulated more Cd in roots and shoots and had significantly lower chlorophyll content, shorter root length, and smaller biomass, suggesting that downregulation of AteIF5A‐2 makes the mutant more Cd sensitive. Real‐time polymerase chain reaction revealed that the expressions of metal transporters involved in Cd uptake and translocation including IRT1, ZIP1, AtNramp3, and AtHMA4 were significantly increased but the expressions of PCS1 and PCS2 related to Cd detoxification were decreased notably in fbr12‐3 compared with Col‐0. As a result, an increase in MDA and H₂O₂ content but decrease in root trolox, glutathione and proline content under Cd stress was observed, indicating that a severer oxidative stress occurs in the mutant. All these results demonstrated for the first time that AteIF5A influences Cd sensitivity by affecting Cd uptake, accumulation, and detoxification in Arabidopsis.     

Bacterial iron detoxification at the molecular level

Iron is an essential micronutrient, and, in the case of bacteria, its availability is commonly a growth-limiting factor. However, correct functioning of cells requires that the labile pool of chelatable “free” iron be tightly regulated. Correct metalation of proteins requiring iron as a cofactor demands that such a readily accessible source of iron exist, but overaccumulation results in an oxidative burden that, if unchecked, would lead to cell death. The toxicity of iron stems from its potential to catalyze formation of reactive oxygen species that, in addition to causing damage to biological molecules, can also lead to the formation of reactive nitrogen species. To avoid iron-mediated oxidative stress, bacteria utilize iron-dependent global regulators to sense the iron status of the cell and regulate the expression of proteins involved in the acquisition, storage, and efflux of iron accordingly. Here, we survey the current understanding of the structure and mechanism of the important members of each of these classes of protein. Diversity in the details of iron homeostasis mechanisms reflect the differing nutritional stresses resulting from the wide variety of ecological niches that bacteria inhabit. However, in this review, we seek to highlight the similarities of iron homeostasis between different bacteria, while acknowledging important variations. In this way, we hope to illustrate how bacteria have evolved common approaches to overcome the dual problems of the insolubility and potential toxicity of iron.     

The effect of manganese oxides and manganese ion on growth and siderophore production by Azotobacter vinelandii

The addition of manganese oxides to iron-limited medium promoted the formation of the pyoverdin siderophore azotobactin by Azotobacter vinelandii. When active-MnO₂ was used, there was greatly decreased iron uptake into the cells, hyperproduction of azotobactin and the abiotic, chemical destruction or adsorbtion of the catechol siderophores azotochelin and aminochelin by this strong oxidizing agent. Although the iron content of the cells was the same as iron-limited cells, the growth of cells in medium with active-MnO₂ was increased 1.5- to 2.5-fold over iron-limited controls. This growth promotion was not caused by iron contaminating the oxide or by manganese solubilized from the oxide. Soluble 0.05–4 mm Mn²⁺ inhibited the growth of iron-limited cells and had a minimal effect on iron uptake, but caused hyperproduction of azotobactin. This later effect was caused by the inhibition of soluble ferric reductase, in a manner identical to that previously observed for Zn²⁺. These results suggest that active-MnO₂ may interfere with a surface-localized iron uptake site, possibly another ferric reductase. The reason for the growth promotion by active-MnO₂ remains unknown, but is most likely related to decreased oxygen toxicity.     

Antinociceptive activity of the volatile oils of Hyptis pectinata L. Poit. (Lamiaceae) genotypes

Hyptis pectinata L. Poit (Lamiaceae) is known popularly in Brazil as “sambacaita” or “canudinho” and is used in the treatment of inflammations, bacterial infections and ache. The antinociceptive activity of the volatile oils of six genotypes, at doses of 100, 200 and 400 mg/kg body wt., were investigated using abdominal writhe models induced by acetic acid and hot plate tests. The volatile oils of all the genotypes are composed mainly of sesquiterpenoids. All the genotypes showed antinociceptive effects in both models used; the SAM002 genotype showed the major inhibitory effect at dose of 100 mg/kg body wt. These results suggest that the volatile oil of H. pectinata has peripheral (writhe reduction) and central (time delay of thermal reaction) effects. These observations indicate that H. pectinata may be useful as an analgesic drug.     

纳米管和镉对蚕豆幼苗根系的毒害效应

以蚕豆幼苗为实验材料,采用水培法研究不同浓度(0、2、4、8、16 mg·L~(-1))羟基化多壁碳纳米管(MWCNTs-OH)与10μmol·L~(-1)镉(Cd)单一和联合作用对蚕豆幼苗根系生长、生理及Cd含量的影响。结果表明:(1)单一MWCNTs-OH处理下,随着MWCNTs-OH浓度(2～8 mg·L~(-1))的增加,蚕豆的根长度增加,且■产生速率增加的同时会伴随着SOD、POD活性升高。(2)单一Cd处理的幼苗根系MDA含量和■产生速率均高于对照,其中■产生速率较对照增加了1.98倍。(3)MWCNTs-OH联合处理诱导了蚕豆根■的产生和MDA含量增加,16 mg·L~(-1) MWCNTs-OH和Cd共同作用对蚕豆幼苗根的损伤最大,会促使部分根尖细胞受损脱落,Cd含量明显下降。研究发现,低浓度MWCNTs-OH可促进蚕豆的根生长,降低Cd的毒性;高浓度MWCNTs-OH(16 mg·L~(-1))和Cd对蚕豆幼苗根的致毒性表现为协同效应。     

Three-year field response of young olive trees (Olea europaea L., cv. Arbequina) to soil salinity: Trunk growth and leaf ion accumulation

Irrigated olive is rapidly increasing in arid and semiarid areas, many of which may be negatively affected by soil salinity. We evaluated changes in trunk growth and leaf Cl^–, Na⁺ and K⁺ concentrations in young Arbequina olives (Olea europaea L.) grown in a saline-sodic field over a three-year period. The trunk diameter was measured at the beginning and the end of the 1999 (70 trees), 2000 (59 trees) and 2001 (42 trees) growing periods. Leaves, sampled in August of each year, were analyzed for Cl^–, Na⁺ and K⁺ concentrations. Soil salinity (apparent electrical conductivity, ECa) of each monitored tree was measured 14 times during the 1999–2001 experimental period with an electromagnetic sensor and converted to root zone electrical conductivity of the soil saturation extract (ECe) based on ECa–ECe calibration curves. Salinity tolerance was determined using the Maas and Hoffman threshold–slope response model. Based on salinity thresholds (ECe_thr), the tolerance of olive in terms of trunk growth was high in 1999 (ECe_thr = 6.7 dS m^–1), but declined with age and time of exposure to salts by 30% in 2000 (ECe_thr = 4.7 dS m^–1) and by 55% in 2001 (ECe_thr = 3.0 dS m^–1). Based on the high absolute slopes obtained in all years (values between 16% and 23% dS^–1 m), olive was classified as very sensitive to ECe values above the threshold. Trunk growth thresholds based on leaf ion concentrations varied, depending on years, between 2.6 and 4.0 mg g^–1 (Cl_thr) and between 1.0 and 1.2 mg g^–1 (Na_thr), indicating that Arbequina olive was less sensitive to leaf Cl^– and much more sensitive to leaf Na⁺ than values reported as toxic in greenhouse studies. Leaf K⁺ slightly decreased with increasing salinity, whereas the K⁺/Na⁺ ratio sharply decreased with increasing salinity. We concluded that the initial salinity tolerance of olive was high, but declined sharply with time of exposure to salts and became quite sensitive due primarily to increasing toxic concentrations of Na⁺ in the leaves.     

Comparative toxicity,growth inhibitory and biochemical effects of terpenes and phenylpropenes on Spodoptera littoralis (Boisd.)

Eleven monoterpenes, phenylpropenes and sesquiterpenes were evaluated for their insecticidal and growth inhibitory activities against the second and fourth larval instars of Spodoptera littoralis. Among the tested compounds, 1,8-cineole revealed the highest fumigant toxicity against the 2nd and 4th larval instars with LC₅₀ values of 2.32 and 3.13 mg/L air, respectively. The monoterpenes, p-cymene, α-terpinene, (−)α-pinene and (−)-carvone were highly toxic to both larval stages as their LC₅₀ values ranged between 7.35 and 13.79 mg/L air against 2nd larval instar and between 14.66 and 32.02 mg/L air against 4th larval instar. In topical application assay against the 4th larval instar, (−)-carvone (LD₅₀ = 0.15 mg/larva) and cuminaldehyde (LD₅₀ = 0.27 mg/larva) were the most potent contact toxicants. In residual film assay, trans-cinnamaldehyde, (−)-citronellal and p-cymene showed the highest insecticidal activity against the 2nd larval instar, while α-terpinene and (−)-carvone were most effective compounds against the 4th larval instar. Moreover, the tested compounds caused strong growth reduction of both larval stages with growth inhibition higher than 80% in the 2nd larval instar and higher than 70% in the 4th larval instar. On the other hand, (−)-carvone, cuminaldehyde and (Z,E)-nerolidol showed pronounced inhibitory effects on acetylcholinesterase (AChE) and adenosine triphosphatases (ATPases) activity of S. littoralis larvae. Cuminaldehyde (IC₅₀ = 1.04 mM) and (Z,E)-nerolidol (IC₅₀ = 0.02 mM) caused the highest inhibition of AChE and ATPases, respectively. Taken together, the results indicate that monoterpenes, phenylpropenes and phenylpropenes could be used to develop new botanical insecticides for S. littoralis management.     

Two-compartment model for rabbit skin organ culture

Summary A new method was developed for rabbit skin organ culture. In a two-compartment model, skin discs were cultured on a Millicell-HA insert unit with a microporous membrane which allows transport of culture medium via the dermis into the epidermis, whereas the epidermal side remains free of direct contact with culture medium. In this relatively simple two-compartment organ culture model, rabbit skin could be cultured for 7 d in RPMI 1640 medium supplemented with fetal bovine serum, or for 2 d in RPMI 1640 medium supplemented with cofactors. The histomorphology and ultrastructure of 7-d cultured rabbit skin discs was essentially similar to that of freshly isolated rabbit skin. Keratinocytes in the stratum basale continued to divide during organ culture. The terminal differentiation of the epidermis continued in vitro as was found by the presence of keratohyalin granules, the intact stratum corneum, and keratin expression. Furthermore, glucose consumption continued until culture Day 7, but thereafter it declined rapidly. Concomitantly, degenerative changes were found. At the end of the 7-d culture period the distance between single dermal collagen fibrils had increased as compared to noncultured skin. This model of skin organ cultures can be used to study biological processes, dermal toxicity, and penetration and metabolism of xenobiotics in intact skin. Furthermore, within certain limits, processes responsible for repair and regeneration of damaged skin can also be studied in this model because the rabbit skin can be cultured for 7 d. The present study was financially supported by grants of Duphar B. V. (Weesp, Netherlands), the European Community, and the Dutch animal welfare organizations Samenwerkingsverband van de Nederlandse Vereniging tot Bescherming van Dieren en de Nederlandse Bond tot Bestrijding van de Vivisectie, Anti-Vivisectie Stichting en Stichting Schoonheid Zonder Wreedheid.     

Challenges in constructing statistically based structure-activity relationship models for developmental toxicity

Regulatory agencies are increasingly called upon to review large numbers of environmental contaminants that have not been characterized for their potential to pose a health risk. Additionally, there is special interest in protecting potentially sensitive subpopulations and identifying developmental toxicants that may be present in the environment. Thus, there is an urgent need for efficient methods to screen large numbers of chemicals for their potential to pose a developmental hazard. One potential screening method involves the use of statistically based structure-activity relationship (SAR) tools to predict activity of untested chemicals. Such systems rely on statistical analyses to discern relationships between structure and activity for a training set of substances. Predictions can then be made for an untested substance as long as its structural features are encompassed by chemicals of the training set. In theory, such systems could assist regulatory agencies in their screening efforts; however, to date, there has been little independent evaluation of these tools for this use. To contribute to such an evaluation, the International Life Sciences Institute Risk Science Institute (ILSI RSI) convened a Working Group to examine methodology used to construct statistically based SAR systems for developmental toxicity. This document reports on the deliberations of the Working Group, which concluded that an improved process is needed for utilizing developmental toxicity data in the construction of statistically based SAR models. The process must be objective, reproducible, rational and transparent. Moreover, it must be informed by the expertise of developmental toxicologists and biologists and must be subject to peer review.