Comparison of distribution of virulence determinants in clinical and environmental isolates of Vibrio cholera

Background: The virulence of a pathogenic Vibrio cholerae is dependent on a discrete set of genetic determinants. In this study, we determined the distribution of virulence determinants among the clinical and environmental isolates of V. cholerae. Methods: The antibiotic resistance profiles of the isolates were determined using standard disk diffusion assay. PCR assay was performed to analyze the presence of toxin genes of ctx, zot and ace. The composition of cholera toxin encoding element (CTX) region flanking of the V. cholerae isolates was also analyzed. Results: All of the clinical isolates (100%) showed a complete set of virulence genes and also the attachment site of the filamentous bacteriophage CTXphi. None of the environmental isolates contained the virulence genes and the attachment site of the CTXphi. Analysis of the flanking regions including the toxin-linked cryptic element and repeat in toxin genes revealed their integrity in the clinical isolates while in the environmental isolates they were absent or contained incomplete sequences. Comparison of the antibiotic resistance assay of the environmental and clinical isolates showed a significant difference in the resistance profiles of the isolates obtained from the two sites. High rates of resistance to co-trimoxosol, streptomycin and chloramphenicol were found with clinical isolates. Conclusion: The absence of all virulence determinants in the environmental strains may suggest that certain ecological features must be present for V. cholerae to acquire a complete set of virulence determinants and to turn them into pathogenic strains.     

Effect of Metasystox-R on marine Nitrosomonas sp. as a nitrification inhibitor

Metasystox-R is a systemic soluble liquid insecticide for the control of aphids on brassica vegetable crops, cotton and lupins and it is possible enter to the marine environment and may be have a hazard effects for the marine organisms and nitrification processes. Effect of Metasystox-R on ammonia oxidizing activity by marine Nitrosomonas sp. was investigated by determining nitrification inhibitor assay in the cell suspension. Results showed that 8 microg mL(-1) of Metasystox-R with PI50 = 4.48 significantly inhibited nitrite production by marine Nitrosomonas sp. These results suggested marine Nitrosomonas sp. may be one of the target bacteria which was inhibitor and decreasing nitrification in the marine environment.     

Computer simulation of the interactions of glyphosate with metal ions in phloem

Essential nutrients such as trace metal ions, amino acids, and sugars are transported in the phloem from leaves to other parts of the plant. The major chelating agents in phloem include nicotianamine, histidine, cysteine, glutamic acid, and citrate. A computer model for the speciation of metal ions in phloem has been used to assess the degree to which the widely used herbicide glyphosate binds to Fe(3+), Fe(2+), Cu(2+), Zn(2+), Mn(2+), Ca(2+), and Mg(2+) in this fluid over the pH range of 8 to 6.5. The calculations show that glyphosate is largely unable to compete effectively with the biological chelating agents in phloem. At a typical phloem pH of 8, 1.5 mM glyphosate binds 8.4% of the total Fe(3+), 3.4% of the total Mn(2+), and 2.3% of the total Mg(2+) but has almost no effect on the speciation of Ca(2+), Cu(2+), Zn(2+), and Fe(2+). As the pH decreases to 6.5, there are some major shifts of the metal ions among the biological chelators, but only modest increases in glyphosate binding to 6% for Fe(2+) and 2% for Zn(2+). The calculations also indicate that over 90% of the glyphosate in phloem is not bound to any metal ion and that none of the metal-glyphosate complexes exceed their solubility limits.     

新疆鼓藻属植物（Cosmarium Cord.）新记录种

乌鲁木齐达坂城地区是新疆湿地分布比较集中的地区之一,包括河流、沼泽、湖泊和池塘等。2010年7-8月,先后2次在该地区采集藻类标本,在对标本鉴定过程中,发现新疆鼓藻属新记录有6种,分别为双眼鼓藻（Cosmarium bioculatum）、扁鼓藻（C.depressum）、丘凸鼓藻（C.ochthodes）、厚皮鼓藻（C.pachydermum）、斑点鼓藻（C.punctulatum）和直角鼓藻（C. rectangulare）,并描述了其形态学特征。这些新记录的发现,丰富了新疆鼓藻属植物的研究资料。     

Modeling of maximum dry density and optimum moisture content of stabilized soil using artificial neural networks

This study considers the use of artificial neural networks (ANNs) to predict the maximum dry density (MDD) and optimum moisture content (OMC) of soil‐stabilizer mix. Multilayer perceptron (MLP), one of the most widely used ANN architectures in the literature, is utilized to construct comprehensive and accurate models relating the MDD and OMC of stabilized soil to the properties of natural soil such as particle‐size distribution, plasticity, linear shrinkage, and the type and quantity of stabilizing additives. Five ANN models are constructed using different combinations of the input parameters. Two separate sets of ANN prediction models, one for MDD and the other for OMC, and also a combined ANN model for multiple outputs are developed using the potentially influential input parameters. Relative‐importance values of various inputs of the models are calculated to determine the significance of each of the predictor variables to MDD and OMC. Inferring the most relevant input parameters based on Garson's algorithm, modified ANN models are separately developed for MDD and OMC. The modified ANN models are utilized to introduce explicit formulations of MDD and OMC. A parametric study is also conducted to evaluate the sensitivity of MDD and OMC due to the variation of the most influencing input parameters. A comprehensive set of data including a wide range of soil types obtained from the previously published stabilization test results is used for training and testing the prediction models. The performance of ANN‐based models is subsequently analyzed and compared in detail. The results demonstrate that the accuracy of the proposed models is satisfactory as compared to the experimental results.     

Exclusion of NEU1 and PPGB from candidate genes for a lysosomal storage disease in Japanese Black cattle

A case of lysosomal storage disease has been reported in a calf of Japanese Black cattle. Lysosomal storage diseases are hereditary diseases caused by deficiency of lysosomal hydrolases. The clinical and pathological features and accumulated substrates of the affected animal indicated a possibility of sialidosis or galactosialidosis caused by deficiency of neuraminidase (NEU1) or protective protein for β-galactosidase (PPGB). In the present study, we investigated nucleotide sequences of the genes encoding these two proteins to evaluate whether mutation of these genes is involved in this disease. We determined cattle genomic sequences of these two genes by using bovine EST sequences and the nucleotide sequences of all exons of these genes were compared between affected and normal animals. The results showed several nucleotide substitutions, but none of them was a functional mutation or specific to the affected animal. Furthermore, genotyping of the microsatellite markers in the vicinity of these two genes revealed no homozygosity of the chromosomal regions including these genes in the affected animal. These findings indicated that neither NEU1 nor PPGB gene is responsible for the lysosomal storage disease of Japanese Black cattle and therefore the disease is neither sialidosis nor galactosialidosis.     

Methylparathion- and carbofuran-induced mitochondrial dysfunction and oxidative stress in Helicoverpa armigera (Noctuidae: Lepidoptera)

The cotton bollworm, Helicoverpa armigera is a polyphagous pest of several crops in Asia, Africa, and the Mediterranean Europe. Organophosphate and carbamate insecticides are used on a large-scale to control Helicoverpa. Therefore, we studied the effect of methylparathion and carbofuran, an organophosphate and carbamate insecticide, respectively, on oxidative phosphorylation and oxidative stress in H. armigera larvae to gain an understanding of the different target sites of these insecticides. It was observed that state III and state IV respiration, respiratory control index (RCI), and P/O ratios were inhibited in a dose-dependent manner by methylparathion and carbofuran under in vitro and in vivo conditions. Methylparathion and carbofuran inhibited complex II by ∼45% and 30%, respectively. Lipid peroxidation, H₂O₂ content, and lactate dehydrogenase (LDH) activity increased and glutathione reductase (GR) activity decreased in a time- and dose-dependent manner in insecticide-fed larvae. However, catalase activity was not affected in insecticide-fed larvae. Larval growth decreased by ∼64% and 67% in larvae fed on diets with 100 μM of methylparathion and carbofuran. The results suggested that both the insecticides impede the mitochondrial respiratory functions and induced lipid peroxidation, H₂O₂, and LDH leak, leading to oxidative stress in cells, which contribute to deleterious effects of these insecticides on the growth of H. armigera larvae, along with their neurotoxic effects.     

Toxico-pathological effects of cypermethrin upon male reproductive system in rabbits

The objective of this work was to identify the reproductive toxico-pathological effects of cypermethrin (CY) in mature male rabbits. Apparently healthy adult age- and weight-matched rabbits (n = 40) were procured from the local market, kept under similar management conditions and divided into four equal groups. Different doses of CY (50, 100 and 150 mg kg⁻¹ body mass) mixed in mustard oil were injected intraperitonealy at weekly intervals in male rabbits (groups B-D) prior to mating. Group A served as control and each animal in this group received equivalent volume of mustard oil. Treatment was continued for 71 days. Decreased testicular and epididymal sperm counts were recorded mostly in dose and time dependent manner in CY-treated rabbits. The serum testosterone concentrations in CY-treated rabbits were significantly lower than those of control group. Degeneration, arrested spermatogenesis and connective tissue (CT) proliferation in testes, while sperm-less seminal plasma and tailless spermatozoa in epididymis of CY-treated rabbits were observed. It was inferred that CY-induced defects in sperms and pathological alterations in testes and epididymis.     

Pharmacokinetics of levamisole after intramuscular and oral administrations to Caspian salmon (Salmo trutta caspius)

The pharmacokinetic parameters of levamisole were determined in the Caspian salmon after intramuscular (IM), oral by gavage, and oral by feed administrations. Eighty-one healthy fish in three different groups received levamisole at the dose of 25 mg/fish by each route. Blood samples were collected at time points of 0, 0.5, 1, 2, 4, 6, 12, 14, and 24 hr after administrations. Plasma levamisole concentrations were measured by a validated high-performance liquid chromatography (HPLC) assay and were analyzed using a noncompartmental approach. The mean terminal half-life was 4.56, 3.95, and 2.91 hr for IM, gavage and feed routes, respectively. The peak plasma concentration for IM, gavage, and feed routes of levamisole were 35.53, 4.63, and 8.36 µg/ml, respectively, at the time of 0.25 for IM, and 1 hr for gavage and feed. The relative bioavailability for gavage and feed routes was 54.80 and 69.30. The similar bioavailability for gavage and feed might be indicative of similar efficacy for these routes of administrations. Further studies are warranted to evaluate the absolute oral bioavailability and the effective dose in Caspian salmon.     

Growth stage-based response of wheat (Triticum aestivum L.) to kinetin under water-deficit environment: pigments and gas exchange attributes

Plant growth regulator, kinetin, is known to modulate the key physiological processes under abiotic stresses in different crops. However, kinetin-mediated response at different growth stages of crop plants is lagging behind. Therefore, a field experiment was conducted to appraise the potential role of exogenously applied kinetin in alleviating the effects of water scarcity on wheat. Three levels of kinetin (0, 75, and 150 mg/L) were used either as seed treatment or foliar spray at the vegetative or the post-anthesis stage. Water deficit markedly reduced shoot fresh mass, plant chlorophyll level, flag leaf photosynthesis, stomatal conductance, and transpiration rate. Degradation of chlorophyll a was greater in plants subjected to post-anthesis water-deficit conditions. However, plants growing under continuous water-deficit conditions had significantly lower concentration of chlorophyll b than those treated with water scarcity at the post-anthesis stage or receiving normal irrigation. Inhibited photosynthesis of wheat in response to post-anthesis water-deficit conditions was largely due to non-stomatal factors. In contrast, stomatal factors were the main constraints for photosynthesis in plants growing under continuous scarcity of water. Plants subjected to continuous water starvation had markedly lower grain yield than those subjected to water-deficit conditions at post-anthesis stage. Application of kinetin before seed sowing or at the post-anthesis stage significantly reduced the negative effects of drought on flag leaf chlorophyll and stomatal conductance. Lower level of kinetin (75 mg/L) was found to be more effective in mitigating the inhibitory effects of water shortage on photosynthesis and growth, and improved grain yield under water scarcity.