Changes in the genetic structure of Bacteria and microbial activity in an agricultural soil amended with tannery sludge

The application of tannery sludge to soils is a form of recycling; however, few studies have examined the impacts of this practice on soil microbial properties. We studied effects of two applications (2006 and 2007) of tannery sludge (with a low chromium content) on the structure of the bacterial community and on the microbial activity of soils. We fertilized an agricultural area in Rolândia, Paraná state, Brazil with different doses of sludge based on total N content, which ranged from 0 to 1200 kg N ha⁻¹. Sludge remained on the soil surface for three months before being plowed. Soils were sampled seven times during the experiment. Bacterial community structure, assessed by denaturing gradient gel electrophoresis (DGGE), was modified by the application of tannery sludge. Soon after the first application, there was clear separation between the bacterial communities in different treatments, such that each dose of sludge was associated with a specific community. These differences remained until 300 days after application and also after the second sludge application, but 666 days after the beginning of the experiment no differences were found in the bacterial communities of the lowest doses and the control. The principal response curve (PRC) analysis showed that the first sludge application strongly stimulated biological activity even 300 days after application. The second application also stimulated activity, but at a lower magnitude and for a shorter time, given that 260 days after the second application there was no difference in biological activity among treatments. PRC also showed that the properties most influenced by the application of tannery sludge were enzymatic activities related to N cycling (asparaginase and urease). The redundancy analysis (RDA) showed that tannery sludge’s influence on microbial activity is mainly related to increases in inorganic N and soil pH. Results showed that changes in the structure of the bacterial community in the studied soils were directly related to changes of their biological activity.     

Changes induced by plastic film mulches on soil temperature and their relevance in growth and fruit yield of pickling cucumber

Purpose: The aim of this study was to realize whether soil mulching, with different plastic mulch colors, is a suitable practice for the culture of pickling cucumber.

Materials and Methods: The crop was cultured or not with black, silver/black, white/black, and aluminum/black plastic films, treatments were evaluated in randomized complete block design, to determine their effect on soil temperature, gas exchange, nutrient concentration, growth, and fruit yield.

Results and Conclusions: Black, silver/black, and aluminum/black plastic mulches were higher (p?≤?0.05) in plant height, leaf area, and shoot dry weight than bare soil at 15 days after sowing, whose results were similar than using white/black film. Maximum, minimum, and mean soil temperatures were higher with all plastic mulches except for aluminum/black, in which the maximum soil temperature was similar to the one in bare soil. There was no difference in net photosynthesis and there was very little difference in nutrient concentration between plants in plastic mulches and plants in bare soil; however, early and total yield showed a higher (p?≤?0.05) yield in all plastic mulches, and lower with bare soil and white/black plastic mulch. Our results confirm that soil mulching impacts the pickling cucumber yield. We suggest a carefull selection of the color plastic mulch.     

Presence of Leptospiral DNA in Semen Suggests Venereal Transmission in Horses

The purpose of the present study was to detect leptospiral DNA by PCR in semen and urine samples of stallions to test for venereal transmission in horses. A total of 10 stallions from four herds were studied, and sampling was conducted in semen and urine for culture and PCR and serum for serology. From the 10 serum samples tested, 6 (60%) were seroreactive. No pure culture was obtained, but leptospiral DNA was detected by PCR in 50% of the semen samples and 30% of urine samples. The present study aimed to detect leptospiral DNA by PCR in semen and urine samples of stallions to test for venereal transmission in horses. Based on these findings, we suggest that there is potential transmission of leptospirosis in horses by sexual transmission.     

High levels of genetic differentiation between Wolbachia-infected and non-infected populations of Folsomia candida (Collembola, Isotomidae)

The Gram-negative -Proteobacterium Wolbachia pipientis has been described as an obligate endosymbiont in many arthropod species, where it induces a variety of reproductive alterations, including parthenogenesis. Recently, this microorganism has also been detected in the parthenogenetic collembolan Folsomia candida. Here, we confirm the occurrence of the endosymbiont also in two Italian parthenogenetic populations of F. candida using ultrastructural (electron microscopy) and molecular (PCR screening on two bacterial genes) evidence. The strain isolated in the Italian populations has almost-identical gene sequences compared to that previously isolated in other populations of F. candida. In addition, we discovered a population of Folsomia cf. candida, which showed the presence of both males and females. This population is not infected by Wolbachia. A screening of two mitochondrial genes (COI and COII) showed that the bisexual population has high levels of genetic divergence in comparison with the parthenogenetic ones, even suggesting the possibility that it belongs to a different species. Furthermore, the remarkably high levels of genetic divergence between the two parthenogenetic populations suggests a possible influence of Wolbachia on inducing such differentiation, and, in the long term, speciation.     

Ecotoxicological evaluation of Harbour sediments using marine organisms from different trophic levels

Background, Aim and Scope  The toxicity of contaminated sediments should be evaluated considering the direct exposure of laboratory organisms to whole sediments and the indirect exposure to elutriates or extracts (Tay et al. 1992, Byrne and Halloran 1999, Nendza 2002). The alga Dunaliella tertiolecta is indicated for the use in toxicity bioassays because it is highly sensitive to several xenobiotics. Harpacticoid copepods have been already used for toxicity testing and Tigriopus fulvus is a promising Mediterranean target-species in ecotoxicology (Todaro et al. 2001, Faraponova et al. 2003, Pane et al. 2005a). In this study, the toxicity of sediments collected in harbour sites of the Northeastern Adriatic Sea was evaluated by growth inhibition test with free living and alginate-immobilized Dunaliella tertiolecta and acute toxicity test with nauplii and adult Tigriopus fulvus with the aim of pointing out the importance to utilize model organisms from different trophic levels in sediment ecotoxicology. Methodology  Elutriates and whole sediments were tested on free living and immobilized (Pane et al. 1998) algal cells, and on laboratory reared copepods. Free-living D. tertiolecta were exposed to diluted elutriates in a static, multi-well plate system. Naalginate immobilized D. tertiolecta were placed in polystyrene inserts fitted with polyester mesh bottoms and exposed to a thin layer (2 mm) of whole sediments in multi-well plates (EPS 1992, Pane and Bertino 1999). Toxicity tests with copepods were carried out on Tigriopus fulvus nauplii (elutriates) and adults (whole sediments and elutriates). Same-aged nauplii useful for toxicity tests were obtained by egg sac detaching and consequent hatching stimulation (Pane et al. 2006). Newborn nauplii (I–II stage) were exposed to elutriates in multi-well plates provided with polystyrene inserts. Adult T. fulvus maintained in polystyrene inserts fitted with polyester mesh bottoms were placed in contact with a thin layer (2 mm) of whole sediment placed on multi-well plate bottoms. All end-points were evaluated after 96 h. Results  In general, the effects increased with the increasing of elutriate concentration up to 50%; the stimulation or inhibition of algal growth was statistically significant in comparison to the control. The inhibiting elutriates induced EC₅₀ variations of algal growth ranging from 66.9% to 74.3%. The mortality of T. fulvus nauplii was always < 25% after treatment with 100% elutriates and < 10% after treatment with 50% dilution; no effect was shown up with 25% dilution; therefore LC₅₀ was not calculable. The effect of elutriates was negligible on adult copepods and LC₅₀ values were never calculable; percent mortality always resulted in < 10% after treatment with whole sediments. Discussion  Both experimental systems gave substantially similar results after exposition to whole sediments and elutriates. During the experiment with algal cells, the immobilization in Na-alginate and the employment of inserts which allowed the contact of organisms with sediments and their easy counting were particularly useful. Likewise, the employment of inserts of adequate mesh size in the tests with copepods allowed the contact of organisms with the sediment and made organism handling and counting easy, as well as the evaluation of mortality. The methodology here described and the utilization of the proposed test-species could have an importance also considering that the current trend in ecotoxicological research is towards finding the most appropriate organism for specific areas of concern by using indigenous species (Mariani et al. 2006) and towards the major significance of chronic and reproductive end-points. Conclusions  Based on the above results, it can be stated that the bioassay with Dunaliella tertiolecta could be a good estimation tool for the ecotoxicological assessment of marine sediments. The immobilization of algae in Na-alginate was seen to be useful to evaluate the toxicity of whole sediments; the employment of polystyrene inserts allowed an improvement of the procedures. T. fulvus nauplii and adults, as other harpacticoids such as Tigriopus japonicus (Yoon et al. 2006), satisfy the basic criteria for the employment of a standard species in marine bioassays. To date only pelagic Acartia tonsa are utilized in the standardized procedure to evaluate the risk assessment of chemicals or wastewaters (ISO 1999). As, on the contrary, the exposure of copepods to solid-phase contaminants it is not yet standardized, the employment of polystyrene inserts improved the procedures for T. fulvus too. So, the rapidity and the possibility to solve practical problems could be the main attractive features of this technique (Pane et al. 2005a) when applied to whole sediments. Recommendations and Perspectives  The methodology here developed being also applicable to long term and reproduction tests should be recommended because it provides relevant information in comparison with other frequently applied, standardized biotests with crustaceans (ISO 1999). The procedure has been shown to be easily applicable to selected marine organisms. ESS-Submission Editor: Prof. Dr. Henner Hollert (henner.hollert@bio5.rwth-aachen.de)     

Simulating in situ ammonia volatilization losses in the North China Plain using a dynamic soil‐crop model

Ammonia (NH₃) volatilization is an important N loss pathway in intensive agriculture of the North China Plain (NCP). Simulation models can help to assess complex N and water processes of agricultural soil–crop systems. Four variations (Var) of a sub‐module for the deterministic, process‐based HERMES model were implemented ranging from simple empirical functions (Var 3 and 4) to process‐oriented approaches (Var 1 and 2) including the main processes of NH₃ volatilization, urea hydrolysis, nitrification from ammonium‐based N fertilizer, and changes in soil solution pH. Ammonia volatilization, plant growth, and changes in ammonium and nitrate pools in the soil over several winter wheat–summer maize double‐crop rotations at three locations in the NCP were simulated. Results were calibrated with two data sets (Dongbeiwang 1, Shunyi) and validated using two data sets (Dongbeiwang 2, Quzhou). They showed that the ammonia volatilization sub‐module of the HERMES model worked well under the climatic and soil conditions of N China. Although the simpler equations, Var 3 and 4, showed lower deviations to observed volatilization across all sites and treatments with a mean absolute error (MAE) of 1.8 and 1.4 in % of applied N, respectively, compared to process‐oriented approaches, Var 1 and 2, with a MAE of 2.2 and 1.9 in % of applied N, respectively. Environmental conditions were reflected better by the process‐oriented approaches. Generally, simulation results were satisfying but simulated changes in topsoil pH need further verification with measurements.     

Seasonal Variations of Sulphate Reduction Rates, Sulphur Pools and Iron Availability in the Sediment of a Dystrophic Lagoon (Sacca di Goro, Italy)

The aim of this work was to analyse factors which regulate sulphide mobility in the sediments of a dystrophic lagoon (Sacca di Goro, the southern lagoon of the Po river delta, Italy). In 1995-96, sediment oxygen demand and variations in organic matter content, redox potential, iron availability, inorganic sulphur concentrations and sulphate reduction rates were measured in sediment profiles at three stations (G, 4 and 17) representative of the main areas of the lagoon. Stations differed mainly in their salinity range and primary producer communities. High concentrations of reactive iron (110-275 µmol ml-1) and low sulphate reduction rates (0.8-16.1 mmol m-2 d-1) were measured in the sediment. Moreover, high concentrations of reactive ferric iron were detected in winter and spring at the stations closest to the freshwater inputs. Nevertheless, in summer, high concentrations of free sulphides were detected in the porewater, although most of the reactive ferrous iron was not sulphide-bound, indicating that not all of the reactive iron pool was available to buffer against sulphide release and thus measures of reactive iron pools may not be a good measure of the true buffering capacity of the sediment. Furthermore, a considerable production of sulphide may occur in the decaying Ulva biomass in the water column, where its concentration will be independent of the potential buffering capacity of iron in the sediment. Therefore when assessing the vulnerability of coastal lagoons to dystrophic events, both the size and availability of the reactive iron pool as well as the site of sulphide production must be taken into consideration.     

Water absorption of freeze-dried meat at different water activities: a multianalytical approach using sorption isotherm, differential scanning calorimetry, and nuclear magnetic resonance

Hydration of freeze-dried chicken breast meat was followed in the water activity range of aw=0.12-0.99 by a multianalytical approach comprising of sorption isotherm, differential scanning calorimetry (DSC), and nuclear magnetic resonance (NMR). The amount of frozen water and the shape of the T2-relaxogram were evaluated at each water content by DSC and NMR, respectively. Data revealed an agreement between sorption isotherm and DSC experiments about the onset of bulk water (aw=0.83-0.86), and NMR detected mobile water starting at aw=0.75. The origin of the short-transverse relaxation time part of the meat NMR signal was also reinvestigated through deuteration experiments and proposed to arise from protons belonging to plasticized matrix structures. It is proved both by D2O experiments and by gravimetry that the extra protons not contributing to the water content in the NMR experiments are about 6.4% of the total proton NMR CPMG signal of meat.     

Quantitation of selected odor-active constituents in dry fermented sausages prepared with different curing salts

The odor-active compounds of dry-fermented sausages with added nitrite or nitrate as curing agents were identified by gas chromatography-olfactometry (GC-O) applying the detection frequency (DF) method. The quantification of these compounds in the sausage was determined by multiple headspace solid-phase microextraction (multiple HS-SPME). There were no specific odor-active compounds related to the use of nitrite or nitrate although there were differences in the DF value of several compounds. The nitrite-added sausages presented higher DF values for ethanol, 1-hexanol, propanoic acid, 2-heptenal, and nonanal while the nitrate-added sausages had higher DF values for phenylacetaldehyde and 3-methyl-butanal. Eighteen compounds were quantified by multiple HS-SPME. Most of them were above their air detection thresholds, but only hexanal, heptanal, and 1-octen-3-ol were in a concentration higher than their oil threshold values. These compounds would probably be the main contributors to the aroma of fermented sausages.     

Vergleichende Untersuchungen der Mineralstoffgehalte geschädigter und nicht geschädigter Rotbuchen (Fagus sylvatica) auf Muschelkalkstandorten im Saarland

Comperative study of the mineral content of damaged and visibly healthy red beeches (Fagus sylvatica) on limestone soils in the Saarland 50-80 years old red beeches from a limestone region in the Saarland, differentiated in apparently healthy and diseased trees, were investigated on their mineral content beginning with the roots over stem up to the leaves. Macro- and microelement contents were different between healthy and diseased trees. Magnesium content in the roots of the latter was markedly reduced. Also zinc and phosphorous were decreased in the damaged trees, while manganese was partially accumulated. On the other side, a high sulphur content was observed in the crown area of both types of trees.