Design,Evaluation and Application of a Continuously Stirred Tank Reactor System for Use in Nitric Acid Air Pollutant Studies

Nitric acid (HNO₃) vapor is a significant component of air pollution. Dry deposition of HNO₃ is thought to be a major contributor to terrestrial loading of anthropogenically-derived nitrogen (N), but many questions remain regarding the physico-chemical process of deposition and the biological responses to accumulation of dry-deposited HNO₃ on surfaces. To examine these processes experimentally, a continuously stirred tank reactor (CSTR) fumigation system has been constructed. This system enables simultaneous fumigation at several concentrations in working volumes 1.3 m dia by 1.3 m ht, allowing for simultaneous fumigation of many experimental units. Evaluation of the system indicates that it is appropriate for long-term exposures of several months duration and capable of mimicking patterns of diurnal atmospheric HNO₃ concentrations representative of areas with different levels of pollution.     

The Effect of Chloride on Yield and Nutrient Interaction in Greenhouse Tomato (Lycopersicon Esculentum Mill.) Grown in Rockwool

The effect of increasing chloride content in nutrient solution on nutrient composition in root environment, interaction of nutrients in leaves and yield of greenhouse tomato cv. ‘Grace F₁’ grown in rockwool were searched. In Experiment I (2004–2005) the levels of 15, 30, 60, and 90 mg Cl·L^?1 but in Experiment II (2006) 30, 60, 90 and 120 mg Cl·L^?1 of nutrient solution were tested. The sources of chloride were water (9.6–10.7 mg Cl·L^?1) and calcium chloride (CaCl₂·2H₂O) but the rest of nutrients and sodium in all treatments were on the same levels. It was found that increasing content of chloride from 15 to 60 mg Cl·L^?1 enhanced the total and marketable fruit yield. Within the range of 60 to 90 mg Cl·L^?1 the yield was on the optimum level but the content of 120 mg Cl·L^?1 declined it. Increasing chloride content in the nutrient solutions was reflected in rising of chlorine content in leaves. The concentration of chloride above 60 mg C·L^?1 reduced the content of nitrogen but above 90 mg C·L^?1 declined the content of calcium, sulfur and zinc in leaves. The antagonism between Cl:N, Cl:Ca: Cl:S and Cl:Zn was appeared. More variable interaction were between Cl:K and Cl:B. At the low levels of chloride, from 15 to 60 mg Cl·L^?1, potassium and boron content were decreased but at the higher ones, from 90 to 120 mg·L^?1, these nutrients had increasing course. It was not found out the effect of chloride contents on macro and microelement contents in nutrient solution emitted from drippers however their content upraising in root medium (rockwool). The highest increase was found out for Na 95.1 and 64.9 % (Exp. I and II - respectively), next for Ca (76.0, 70.1 %), Cu (62.5 and 71.0 %), Cl (43.6, 24.4), B (33.3, 21.0 %), N-NO₃ (30.4, 49.6 %), Zn (29.5, 32.8 %), S-SO₄ (25.9, 25.5 %), K (24.5, 24.1 %), Fe (19.8, 19.2 %), Mn (17.5, 21.3 %) and Mg (14.9, 11.7). Advantageous effect of chloride on tomato yield justified the need to introduce for the practice adequate chlorine nutrition, and recommend to maintain 60 to 90 mg Cl·L^?1 in nutrient solution. The best yield appeared when content of chlorine in leaves (8th or 9th leaf from the top) was in the range 0.48-0.60 % of Cl in d. m.     

Interaction between protein, phytate, and microbial phytase. In vitro studies

The interaction between protein and phytate was investigated in vitro using proteins extracted from five common feedstuffs and from casein. The appearance of naturally present soluble protein-phytate complexes in the feedstuffs, the formation of complexes at different pHs, and the degradation of these complexes by pepsin and/or phytase were studied. Complexes of soluble proteins and phytate in the extracts appeared in small amounts only, with the possible exception of rice pollards. Most proteins dissolved almost completely at pH 2, but not after addition of phytate. Phytase prevented precipitation of protein with phytate. Pepsin could release protein from a precipitate, but the rate of release was increased by phytase. Protein was released faster from a protein-phytate complex when phytase was added, but phytase did not hydrolyze protein. Protein was released from the complex and degraded when both pepsin and phytase were added. It appears that protein-phytate complexes are mainly formed at low pH, as occurs in the stomach of animals. Phytase prevented the formation of the complexes and aided in dissolving them at a faster rate. This might positively affect protein digestibility in animals.     

Evaluation and Modification of the OECD 301F Respirometry Biodegradation Test Method with Regard to Test Substance Concentration and Inoculum

The majority of OECD (Organisation for Economic Co-operation and Development) ready biodegradation test methods require a relatively high concentration of test substance. Such high dosage concentrations are unsuitable for screening of inhibitory or biocidal compounds. Unfortunately, the method with the lowest dosage concentration (Closed Bottle Test, 301D) also has the lowest biodegradation potential. Therefore, we have evaluated the ability of the Manometric Respirometry (301F) test method to study biodegradation of test substances at concentrations of a few mgl^?1, using sodium acetate as a model substance. Results have been compared to those obtained from closed bottle tests (301D). Problems with using activated sewage as an inoculum have been demonstrated, as respirometry curves were biphasic, and the recorded biological oxygen demand (BOD) was higher than the calculated theoretical oxygen demand (ThOD). A consortium of bacteria isolated from activated sewage was used as an alternative inoculum. Using the alternative inoculum for respirometry tests, monophasic growth curves were observed with recorded BOD values at expected levels. Sensitivity thresholds for the respirometer tests were comparable to those in closed bottles, with biodegradation being detected at concentrations as low as 2.5 mgl^-1 ThOD. Comparison of whole community carbon source utilisation profiles for both inocula (obtained using BIOLOG microplates) revealed their catabolic potential to be similar. Therefore, use of an activated sewage-derived inoculum is recommended as an alternative inoculum for respirometer-based biodegradation testing.     

Correlation between earthworms and plant litter decomposition in a tropical wet forest of Puerto Rico

Earthworms are recognized to play an important role in the decomposition of organic materials. To test the use of earthworms as an indicator of plant litter decomposition, we examined the abundance and biomass of earthworms in relation to plant litter decomposition in a tropical wet forest of Puerto Rico. We collected earthworms at 0–0.1 m and 0.1–0.25 m soil depths from upland and riparian sites that represent the natural variation in soils and decomposition rates within the forest. Earthworms were hand-sorted and weighed for both fresh and dry biomass. Earthworms were dominated by the exotic endogeic species Pontoscolex corethrurus Müller; they were more abundant, and had higher biomasses in the upland than in riparian sites of the forest. Plant leaf litter decomposed faster in the upland than riparian sites. We found that earthworm abundance in the upper 0.1 m of the soil profile positively correlated with decomposition rate of plant leaf litter. Ground litter removal had no effect on the abundance or biomass of endogeic earthworms. Our data suggest that earthworms can be used to predict decomposition rates of plant litter in the tropical wet forest, and that the decomposition of aboveground plant litter has little influence on the abundance and biomass of endogeic earthworms.     

Mercury content and its bioconcentration factors in wild mushrooms at Łukta and Morag,northeastern Poland

Total concentrations of mercury were determined using cold-vapor atomic absorption spectroscopy (CV-AAS) in the fruiting bodies of 16 species of wild mushrooms and underlying soil (0-10 cm) substrates collected in the areas of the Communes of Morag and ?ukta in the county of Ostróda in northeastern Poland in 1997-1998. A total of 174 composite samples of caps, 174 stalks, 80 whole fruiting bodies (collectively 1254 specimens), and 252 soils were examined. Among several species of mushrooms analyzed, the greatest concentrations were between 1300 and 71000 ng.g(-1) of dry matter. These levels were found in the caps of Sweating mushroom (Clitocybe rivulosa), King Bolete (Boletus edulis), and Common Puffball (Lycoperdon perlatum) and also were characterized by the highest bioconcentration factors (BCF) for Hg, which ranged between 160 +/- 82 and 110 +/- 34. The cap to stalk quotient for mercury concentrations was approximately 2 for most of the species except Poison Pax (Paxilus involutus), which had a greater concentration in caps than in stalks and a quotient of 4.4 +/- 7.2. Hg concentrations in the underlying soil substrates (0-10 cm layer) ranged between 21 +/- 21 and 390 +/- 130 ng.g(-1) of dry matter. The results showed that the consumption of mushrooms, considered to be the sole dietary source of mercury at the highest or mean element concentrations found, is not hazardous at daily ingestion rates of less than 70 and 210 g of fresh product, which would result in a hazard index value of less than unity.     

Development of a rapid method for the sequential extraction and subsequent quantification of fatty acids and sugars from avocado mesocarp tissue

Methods devised for oil extraction from avocado (Persea americana Mill.) mesocarp (e.g., Soxhlet) are usually lengthy and require operation at high temperature. Moreover, methods for extracting sugars from avocado tissue (e.g., 80% ethanol, v/v) do not allow for lipids to be easily measured from the same sample. This study describes a new simple method that enabled sequential extraction and subsequent quantification of both fatty acids and sugars from the same avocado mesocarp tissue sample. Freeze-dried mesocarp samples of avocado cv. Hass fruit of different ripening stages were extracted by homogenization with hexane and the oil extracts quantified for fatty acid composition by GC. The resulting filter residues were readily usable for sugar extraction with methanol (62.5%, v/v). For comparison, oil was also extracted using the standard Soxhlet technique and the resulting thimble residue extracted for sugars as before. An additional experiment was carried out whereby filter residues were also extracted using ethanol. Average oil yield using the Soxhlet technique was significantly (P < 0.05) higher than that obtained by homogenization with hexane, although the difference remained very slight, and fatty acid profiles of the oil extracts following both methods were very similar. Oil recovery improved with increasing ripeness of the fruit with minor differences observed in the fatty acid composition during postharvest ripening. After lipid removal, methanolic extraction was superior in recovering sucrose and perseitol as compared to 80% ethanol (v/v), whereas mannoheptulose recovery was not affected by solvent used. The method presented has the benefits of shorter extraction time, lower extraction temperature, and reduced amount of solvent and can be used for sequential extraction of fatty acids and sugars from the same sample.     

Using BCG,MPT-51 and Ag85 as antigens in an indirect ELISA for the diagnosis of bovine tuberculosis

This study evaluated the Mycobacterium tuberculosis protein antigen MPT-51, the trimeric antigen 85 (Ag85) complex, and Bacillus Calmette-Guérin (BCG) in an indirect ELISA to diagnose bovine tuberculosis (TB) from serum samples. Serum was collected from 208 intra-dermal tuberculin test (ITT)-positive and 54 ITT-negative animals from a region where bovine TB is endemic. Using the Ag85 and BCG antigens, the indirect ELISA was able to discriminate ITT-positive from ITT-negative animals. This level of discrimination was not achieved when using the MPT-51 antigen. The highest sensitivity (Se) and specificity (Sp) of the test was found when BCG was used (Se, 82%; Sp, 91%). Further work in different epidemiological settings and with larger numbers of animals will be required to validate these findings.     

Nature conservation in Spain: A brief account

The history of nature conservation in Spain is reviewed with reference to public attitudes past, present and future. The eight National Parks are listed and described in terns of their location, date of foundation, extension and main historical, floristic and faunistic characteristics.The various other types of reserve and their aims and functions are alluded to and the significance of the role played by ICONA (National Institute for the Conservation of Nature) through the declaration and administration of these vast areas is stressed.Ample comment is made on the deficiency of past and current legislation and hope is expressed in connection with the May 1975 Law for the Creation and Conservation of Protected Natural Spaces.     

A rapid chronometric assay of peroxidase activity in citrus leaf discs

A rapid peroxidase assay is proposed, using 3-mm diameter leaf discs, and based on the chronometric measurement of the oxidation of ascorbic acid, which is detected by means of the appearance of the blue colour of the benzidine-coupled reaction.The proposed method is adequately reproducible and sensitive for the diagnosis of iron deficiency in citrus.