Characterization and Inhibitor Studies of Chitinases from a Parasitic Blowfly (Lucilia cuprina), a Tick (Boophilus microplus), an Intestinal Nematode (Haemonchus contortus) and a Bean (Phaseolus vulgaris)

The molecular weight pattern and the stage-specific activities of chitinases from the blowfly Lucilia cuprina, the tick Boophilus microplus and the intestinal nematode Haemonchus contortus were examined. Chitinolytic enzymes could be detected in all parasite species tested, but the activity was different between the stages. Highest chitinolytic titers were found in blowfly pupae (83 kDa, 118 kDa), hatching larvae of ticks (58 kDa, 94 kDa) and nematode eggs (43 kDa). Leaves from ethylene-treated bean plants Phaseolus vulgaris expressed two basic Class I chitinases (Ia, Ib) of 34 kDa, differing in their amino acid sequences at residue 33 and 34 (Ia: glycine, proline; Ib: lysine, aspartic acid). Inhibitor studies with blowfly pupae revealed that allosamidin (IC₅₀=0·32 (±0·02) μM ) was by far the best inhibitor when compared with various amino sugar derivatives. This compound also inhibited chitinases from tick larvae (IC₅₀=0·69(±0·10) μM ) and nematode eggs (IC₅₀=0·048(±0·0045) μM ) specifically. Whereas Class Ia chitinase from bean leaves was inhibited only up to 18% by 10 μM allosamidin, it had an IC₅₀ of 1(±0·14) μM for the Ib type, which is the first plant chitinase described to be highly sensitive to allosamidin.     

Experimental determinations of factors affecting the sink rates of baited hooks to minimize seabird mortality in pelagic longline fisheries

• 1. An experiment was conducted in Australia's pelagic longline fishery to establish a scientific basis for the introduction of line weighting to reduce seabird mortality. The experiment examined the effects of different bait species (blue mackerel, yellow‐tail mackerel and squid), bait life status (dead or alive), weight of leaded swivels (60 g, 100 g and 160 g) and leader length (distance between leaded swivel and hooks: 2 m, 3 m and 4 m) on the sink rates of baited hooks from 0–6 m deep.
• 2. On average, live bait sank much more slowly than dead bait. The sink rates of individual live bait were highly variable: many were <2 m underwater 18 s after deployment, including some on the heaviest swivels, and some were <10 m deep after 120 s.
• 3. Within the dead bait group, all three swivel weights on 3 m and 4 m leaders sank at similar rates. Initial sink rates (e.g. 0–2 m) were 2–3 times slower than final rates (e.g. 4–6 m) for all combinations of swivel weight and leader length. The fastest initial and final sink rates were associated with heavy swivels placed close to hooks.
• 4. The results show that (a) compared with dead bait, live bait greatly increases the exposure of baited hooks to seabirds; (b) initial sink rates of dead bait are increased by placing leaded swivels close to hooks and final rates by increasing the weight of the swivels; (c) adding weight to long leaders makes little difference to sink rates; and (d) the small (incremental) changes to swivel weights and leader lengths typically preferred by industry will be difficult to detect at sea and unlikely to substantially reduce seabird mortality.
• 5. We suggest that experiments designed to reduce seabird mortality from that associated with 60 g swivels and ～3.5 m leaders (the preferred option by industry) should aim to expedite the initial sink rates as well as rates to deeper depths. This objective could be achieved by including branch lines with ≥120 g swivels ≤2 m in comparative assessments of the effectiveness of line weighting regimes in reducing seabird mortality. Copyright © 2010 John Wiley & Sons, Ltd.

     

Tuna viscera protein hydrolysate: nutritive and disease resistance properties for Persian sturgeon (Acipenser persicus L.) larvae

The effect of fish protein hydrolysate (FPH) on the growth, feed utilization, digestive enzymes, gut microflora and resistance to bacterial challenge in Persian sturgeon (Acipenser persicus) larvae during a 45‐day feeding trial has been investigated. Artificial diets containing different levels of tuna visceral protein hydrolysate (PH0, PH10, PH25 and PH50) and live foods (Artemia: 5 days, Daphnia: 40 days) were fed to Persian sturgeon larvae. Larvae fed on the PH10 and PH25 had growth, feed utilization, protein content and trypsin activity significantly better than PH0 and PH50 groups (P < 0.05). Condition factors, survival rate and pepsin activity were not significantly different (P > 0.05). Brush border membrane enzymes to cytosolic enzyme ratio revealed that maturation had been occurred in 41 days post hatch (dph) at moderate FPH levels. In 54 dph larvae, this ratio was not affected by the level of FPH in the diet. No significant differences were observed in the total number of gut microflora or in the resistance to Aeromonas hydrophila (P > 0.05), whereas lactic acid bacteria was significantly higher in PH50 (P < 0.05). The results of this study show that application of moderate levels of FPH in Persian sturgeon diets could enhance larvae performance due to the early maturation in intestinal.     

Combining agent-based and stock-flow modelling approaches in a participative analysis of the integrated land system in Reichraming, Austria

The integrated modelling of coupled socio-ecological systems in land-change science requires innovative model concepts capable of grasping the interrelations between socioeconomic and natural components. Here, we discuss the integrated socio-ecological model SERD (Simulation of Ecological Compatibility of Regional Development) that was developed for the municipality of Reichraming in Upper Austria in a participative 2-year process involving local stakeholders. SERD includes three main components: (1) an agent-based actors module that simulates decisions of farmsteads, the municipal administration and other important actors; (2) a spatially explicit (GIS based) land-use module that simulates land-use change at the level of individual parcels of land and (3) an integrated socio-ecological stock-flow module that simulates carbon and nitrogen flows through both socioeconomic and ecological system compartments. We report on outcomes of a scenario analysis that outlines possible future trajectories depending on both external (e.g. agricultural subsidies and prices) and internal (e.g. innovation, willingness to co-operate) factors. We find that both external and internal factors can affect the behaviour of the integrated system considerably. Local and regional policies are found to be able to counteract adverse global socioeconomic conditions to some extent, but not to reverse the trend altogether. We also find strong interdependencies between socioeconomic and ecological components of the system. Fully evaluating these interdependencies is, however, not possible at the local scale alone and will require explicit consideration of higher-level effects in future research.     

Conserving indigenous crayfish: stock assessment and habitat requirements in the threatened Austropotamobius italicus

• 1. As part of the Austropotamobius pallipes species complex, the crayfish Austropotamobius italicus is a species of community interest whose preservation requires the designation of Special Areas of Conservation (SACs) (Annex II, EU Habitats Directive). This study aimed at (1) assessing the conservation status of this threatened indigenous species by stock assessment in central Italy and (2) identifying some aspects of its elective habitat.
• 2. Surveys were conducted in nine streams harbouring A. italicus (streams WI) and in 10 streams where crayfish populations became extinct at least 5 years before the study (streams WO).
• 3. The results confirmed that A. italicus is a K‐selected species, with a relatively slow growth rate (males: 0.34; females: 0.37) and a long life expectancy (males: 8.2 years; females: 7.8 years). The extant populations are healthy, showing balanced sex‐ratios and well structured age‐class compositions. Mortality is mainly due to fishing, which is illegal in Tuscany.
• 4. Principal Components Analyses showed that the streams WI and WO differ in the abundance of allochthonous plant detritus but not in the taxonomic composition of their macroinvertebrate communities. Age classes were found to be spatially segregated, juveniles mainly using cobbles as substrates and adults seemingly avoiding them.
• 5. The loss of the pristine riverine landscape seems to have been responsible, together with illegal fishing, for the local extinction of the species. As a consequence, retaining, enhancing, and restoring the habitat and its complexity are required for the preservation of A. italicus.
• 6. The designation of SACs might help in this endeavour if accompanied by programmes aimed at publicizing the need for conservation of this species. Unfortunately, crayfish‐focused projects supported by LIFE in Italy since 1992 (4%) and the SACs involved (1.4%) have been relatively few, despite the poor conservation status of this species and its well recognized ecological role.

Copyright © 2008 John Wiley & Sons, Ltd.     

Odor thresholds of microbially induced off-flavor compounds in apple juice

Microbially derived off-flavor is a major problem in apple juice production as it diminishes the sensory quality of the juice significantly. Fifteen relevant off-flavor compounds that are formed in apple juice, for example, by the strains Alicyclobacillus acidoterrestris and Actinomycetes (Streptomyces ssp.) were investigated with respect to their sensory relevance. The odor threshold values (i.e., detection and recognition values) were determined for all compounds in the matrix apple juice. Odor threshold values for fenchyl alcohol are reported here for the first time. The obtained values were set in relation to the limits of detection and quantification of a previously published GC-MS method. Eight tainted apple juice samples were analyzed for the presence of the 2 strains and the 15 off-flavor compounds. Both strains could be found in the samples; the presence of Streptomyces ssp. as spoilage bacteria of apple juice is reported for the first time. In samples with distinct off-flavor, 2-isopropyl-3-methoxypyrazine, 2-isobutyl-3-methoxypyrazine, 2-methylisoborneol, 1-octen-3-ol, fenchyl alcohol, geosmin, and guaiacol as well as 2,6-dibromophenol were determined in concentrations higher than the detection threshold.     

Residue level, persistence, and storage performance of citrus fruit treated with fludioxonil

The potential of postharvest dip treatments with fludioxonil (FLU) (a synthetic analogue of the bacterial metabolite of pyrrolnitrin), in controlling postharvest decay caused by Penicillium digitatum and Penicillium italicum of citrus fruit was investigated in comparison with the conventional fungicide imazalil (IMZ). The ultrastructural changes of fruit epicuticular wax was investigated as a function of water dip temperature, and the possible role of these changes was related to residue accumulation under FLU treatment. Residues retained by fruit were determined as a function of fungicide concentration, dip temperature, and fruit storage conditions. Scanning electron microscopy analysis revealed that fruit dipping in water at 30 or 40 degrees C did not cause differences in cuticular wax's ultrastructure in comparison to control fruit, while treatments at 50, 55, or 60 degrees C caused the disappearance of wax platelets, resulting in relatively homogeneous skin surface, due to partial "melting" of epicuticular wax. Residues of FLU in fruit treated at 20 or 50 degrees C were significantly correlated with the doses of fungicide applied. When equal amounts of fungicide were employed, the residue concentrations were notably higher (from 2.6- to 4-fold) in fruit treated at 50 degrees C than in fruit treated at 20 degrees C. The dissipation rate of FLU in "Salustiana" and "Tarocco" oranges was lower in fruit subjected to treatment at 50 degrees C. The minimal FLU concentration for almost complete decay control in artificially wounded fruit during 7-d storage at 20 degrees C was 400 mg/L active ingredient (ai) in fruit treated at 20 degrees C and 100 mg/L ai in fruit treated at 50 degrees C. Results on nonwounded Tarocco oranges subjected to 3 weeks of simulated quarantine conditions at 1 degrees C, plus 6 weeks of standard storage at 8 degrees C and an additional two weeks of simulated marketing period (SMP) at 20 degrees C revealed that almost complete decay control with FLU applications of 100 mg/L at 50 degrees C and 400 mg/L at 20 degrees C resulted in ca. 0.8 mg/kg FLU fruit residues, in agreement with results on wounded citrus fruit. When equal concentrations and temperatures were applied, FLU treatments were as effective as IMZ. In vitro trials showed a low sensitivity to FLU against P. digitatum and P. italicum isolates. MIC values for the complete inhibition of mycelium growth were >or=100 microg/mL, while ED(50) values ranged from 0.1 to 1 microg/mL for P. digitatum and from 1 to >100 microg/mL for P. italicum. The latter result suggests that care should be taken to avoid exclusive application of FLU in a sustainable program for management of fruit decay. However, integrating fungicide application and hot water dip may reduce the possibility of selecting fungicide-resistant populations of the pathogen, by increasing the effectiveness of the treatment.     

Synthesis and phytotoxicity of new ionic liquids incorporating chiral cations and/or chiral anions

The aim of this work was to synthesize chiral ionic liquids as chiral solvents for organic synthesis and to evaluate the phyto(eco)toxicity of the new products and starting N-alkylimidazoles and their potential environmental influence on soil and plants. Chiral ionic liquids containing anions such as Cl-, Br-, TsO-, PF6(-), NO3(-), CF3SO3(-), and (+)- and (-)-C6H5CH(OH)C(O)O- were synthesized using (-)-(1R)-6,6-dimethylbicyclo[3.1.1]hept-2-ene-2-ethyl [(-)-(1R)-nopyl] halides (X = Cl, Br) and tosylate in 62-100% yields. The chloride 7 and the nitrate 13 ionic liquids possessed a toxicity dependent on the applied concentration. The lowest concentration causing a distinct reduction in plant germination/growth was 100 mg/kg. Spring barley better tolerated the ionic liquids (200 mg/kg) than common radish (100 mg/kg). The nitrate liquid did not exhibit an inhibiting effect on the germination ability of seeds. The starting N-methylimidazole used in lower concentrations (1 and 10 mg/kg of soil dry weight) was not phytotoxic, in contrast to higher doses (>1000 mg/kg).     

CD46 in meningococcal disease

The human-specific bacterial pathogen Neisseria meningitidis is a major cause of sepsis and/or meningitis. The pili of N. meningitidis interact with CD46, a human cell-surface protein involved in regulation of complement activation. Transgenic mice expressing human CD46 were susceptible to meningococcal disease, because bacteria crossed the blood-brain barrier in these mice. Development of disease was more efficient with piliated bacteria after intranasal, but not intraperitoneal, challenge of CD46 transgenic mice, suggesting that human CD46 facilitates pilus-dependent interactions at the epithelial mucosa. Hence, the human CD46 transgenic mice model is a potentially useful tool for studying pathogenesis and for vaccine development against meningococcal disease.