Recovery of Francisella tularensis from soil samples by filtration and detection by real-time PCR and cELISA

The aim of this study was to develop a specific and highly sensitive method able to detect very low concentrations of Francisella tularensis in soil samples by real-time PCR (qPCR) with SYBR Green I. tul4 gene, which encodes the 17-kDa protein (TUL4) in F. tularensis strains, was amplified using a LightCycler (LC) device. We achieved a detection limit of 0.69 fg of genomic DNA from F. tularensis subp. holarctica live vaccine strain (LVS), corresponding to a value less than 3.4 genome equivalents per reaction. The qPCR was shown to be specific, highly sensitive and reproducible. In addition, we evaluated 2 new methods for recovering bacteria from soil based on 1-step filtration using glass fiber filters and PVDF filters. These filtration methods enabled us to recover F. tularensis efficiently from soil samples. As few as 50 CFU per 0.5 g of soil were detected by qPCR. Capture enzyme-linked immunosorbent assay (cELISA) allowed us to detect and quantify the amount of bacteria recovered from soil by an immunological method. Although qPCR was more sensitive than cELISA, we did not observe substantial differences in the amount of bacteria quantified by both methods.     

Effect of liquid cow manure on andisol properties and atrazine adsorption

Application of animal manure amendments to agricultural soils is a common practice to improve soil fertility through the addition of essential plant nutrients. This practice may increase the potential for atrazine (2-chloro-4-ethylamino-6-isopropylamino-1, 3, 5-triazine) leaching due to competition for adsorption sites between the pesticide and dissolved organic carbon (DOC) added through manure. We evaluated the influence of liquid cow manure (LCM) application on soil properties, atrazine adsorption, and the physicochemical controlling mechanisms in an Andisol. The LCM was applied at rates equivalent to 0, 100,000, 200,000, and 300,000 L ha(-1), resulting in treatments S-0, S-100, S-200, and S-300, respectively. The LCM application increased DOC and pH of the soils immediately on addition, but pH returned to S-0 values 30 d after application. The LCM application did not modify atrazine adsorption with the two lowest application rates (S-100 and S-200), but atrazine adsorption was decreased in S-300 (K(f) = 0.96) compared with the control (S-0) (K(f) = 1.19), possibly due to the competitive adsorption of DOC with the pesticide. The Fourier-transformed infrared analysis showed that LCM increased aliphaticity and presence of N-containing groups and polysaccharide-like groups in amended soils; however, these properties did not modify the atrazine interaction in the studied amended soils. Interestingly the addition of DOC to soil at the high application rate (S-300) reduced atrazine adsorption in this rich OM Andisol despite the LCM not raising the concentration of stable organic matter. The application of high rates of liquid manure containing DOC incurs an increased risk of pesticide leaching.     

Recreation demand analysis under truncation, overdispersion, and endogenous stratification: an application to Gros Morne National Park

Using on-site survey data from Gros Morne National Park in Newfoundland, this paper estimates and compares several truncated count data models of recreation demand. The selected model accounts not only for the truncated and overdispersed nature of the data but also for endogenous stratification due to the oversampling of avid users. Moreover, the model that allows for a flexible specification of the overdispersion parameter dominates on the basis of goodness of fit. The results are used to estimate the users' value of access to the park.     

High-impact conservation: invasive mammal eradications from the islands of western México

Islands harbor a disproportionate amount of the earth's biodiversity, but a significant portion has been lost due in large part to the impacts of invasive mammals. Fortunately, invasive mammals can be routinely removed from islands, providing a powerful tool to prevent extinctions and restore ecosystems. Given that invasive mammals are still present on more than 80% of the world's major islands groups and remain a premier threat to the earth's biodiversity, it is important to disseminate replicable, scaleable models to eradicate invasive mammals from islands. We report on a successful model from western México during the past decade. A collaborative effort between nongovernmental organizations, academic biologists, Mexican government agencies, and local individuals has resulted in major restoration efforts in three island archipelagos. Forty-two populations of invasive mammals have been eradicated from 26 islands. For a cost of USD 21,615 per colony and USD 49,370 per taxon, 201 seabird colonies and 88 endemic terrestrial taxa have been protected, respectively. These conservation successes are a result of an operational model with three main components: i) a tri-national collaboration that integrates research, prioritization, financing, public education, policy work, capacity building, conservation action, monitoring, and evaluation; ii) proactive and dedicated natural resource management agencies; and iii) effective partnerships with academic researchers in Mexico and the United States. What is now needed is a detailed plan to eradicate invasive mammals from the remaining islands in the region that integrates the needed additional financing, capacity, technical advances, and policy issues. Island conservation in western Mexico provides an effective approach that can be readily applied to other archipelagos where conservation efforts have been limited.     

Combining TiO2-photocatalysis and wetland reactors for the efficient treatment of pesticides

In the present work the photocatalytic and biological degradation of two commercial mixtures of pesticides (Folimat and Ronstar) and two fungicides (pyrimethanil and triadimenol) has been studied. The evolution of some components of these commercial products (dicofol, tetradifon and oxadiazon) and that of the two fungicides has been monitored by means of HPLC, GC-MS, TOC and toxicity (Lemna minor toxicity test) measurements. The photocatalytic method was able to degrade dicofol, tetradifon, pyrimethanil, triadimenol and the components of Ronstar with the exception of oxadiazon. In addition to this, the photocatalytic method eliminated pyrimethanil toxicity and reduced that of triadimenol by a 90%, Ronstar by a 78% and Folimat by an 87%. Nevertheless, the wetland reactors alone could reduce the toxicity of only the former. Finally, the proper dosage of the water containing the pesticides to a photocatalytic reactor followed by a wetland reactor resulted to be the most successful strategy for the detoxification of the studied compounds and their mixtures.     

Evaluation of the Spanish hot dip galvanising sector as a source of polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans

A survey to estimate the polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) emissions of Spanish hot dip galvanising sector was carried out during 2002. This investigation is the first presenting Spanish experimental data related to this industrial sector. Three different matrices: flue gas, ash and filter dust were tested to quantify the PCDD/Fs generated during the galvanising process. The organic source of PCDD/F formation could be from the insufficient degreasing o from inhibitors or additives used in the pickling steps such as quinoline, isoquinoline, 8-methylquinoline or polyether phosphoric acid. Low levels PCDD/Fs were achieved in air emissions when air control devices are used. On the contrary, filter dusts are highly contaminated; indicating that the absence of air control devices would increase the risk of fugitive emissions. Homologue profiles and Principal Component Analysis demonstrate there are differences in the formation mechanisms in the bath zone (ashes) compared to the stack location (filter dusts and air emissions), related to the de novo synthesis and reaction time. The annual PCDD/F emission to the atmosphere for this sector during 2002 has been estimated in 0.023g I-TEQ. The emission factor of plants with air control devices has been calculated at 0.030microg I-TEQ/ton of galvanised steel.     

Solar TiO2-assisted photocatalytic degradation of IGCC power station effluents using a Fresnel lens

The heterogeneous TiO2 assisted photocatalytic degradation of wastewater from a thermoelectric power station under concentrated solar light irradiation using a Fresnel lens has been studied. The efficiency of photocatalytic degradation was determined from the analysis of cyanide and formate removal. Firstly, the influence of the initial concentration of H2O2 and TiO2 on the degradation kinetics of cyanides and formates was studied based on a factorial experimental design. Experimental kinetic constants were fitted using neural networks. Results showed that the photocatalytic process was effective for cyanides destruction (mainly following a molecular mechanism), whereas most of formates (degraded mainly via a radical path) remained unaffected. Finally, to improve formates degradation, the effect of lowering pH on their degradation rate was evaluated after complete cyanide destruction. The photooxidation efficiency of formates reaches a maximum at pH around 5-6. Above pH 6, formate anion is subjected to electrostatic repulsion with the negative surface of TiO2. At pH<4.5, formate adsorption and photon absorption are reduced due to some catalyst agglomeration.     

Application of fluorescence in situ hybridization technique to detect simazine-degrading bacteria in soil samples

We propose a new approach to evaluate the natural attenuation capacity of soil by using fluorescence in situ hybridization (FISH). A specific oligonucleotide probe AtzB1 was designed based on the sequence data of the atzB gene involved in the hydrolytic deamination of s-triazines; this gene, located in a multiple copy plasmid was detected by the optimized FISH protocol. Two agricultural soils (Lodi and Henares) with a history of simazine treatments, and two natural soils (Soto and Monza), without previous exposure to simazine, were studied. AtzB1 probe-target cells were found only in the agricultural soils and, in a greater percentage, in the Lodi soil, compared to the Henares one. Moreover, the greatest percentage of AtzB1 probe-target cells in Lodi was accompanied by a greater mineralization rate, compared to the Henares soil. The FISH method used in this study was suitable for the detection of simazine-degrading bacteria and could be a useful indicator of the potential of soil bioremediation.     

Evaluation of carbon degradation during co-composting of exhausted grape marc with different biowastes

In this work the carbon biodegradation of exhausted grape marc (EGM) combined with other organic wastes using the turned pile composting system was studied. Four different piles were made of EGM in Pile 1, EGM mixed with cow manure and straw (CMS) in Pile 2, EGM mixed with municipal solid waste (MSW) in Pile 3 and EGM mixed with grape stalks (GS) in Pile 4. The results obtained were modelled to determine the main kinetic and stoichiometric parameters. Regarding to the rate constants of the composting processes they were increased from 0.033d(-1), the value obtained when EGM was composted alone, to 0.040 and 0.044d(-1) when MSW and GS were added, respectively as co-substrates. However, the addition of CMS reduced the rate constant. About the biodegradable carbon fractions, it was observed that the co-composting reduced significantly the remanent carbon concentration after composting in all the piles whilst increased the readily biodegradable carbon fractions from 35, the value obtained when EGM was composted alone, to 50 and 60%, respectively when MSW or GS were added. As regards the temperature profiles, only Piles 1 and 4 achieved thermal hygienization values and about the nitrogen losses, the lowest percentage of nitrogen loss took place when GS were added, because of its optimum pH and C/N initial ratio. Thus, though any of these wastes could be used for co-composting with EGM, the use of GS as co-substrate and bulking agent for the co-composting process of EGM was recommended.     

Effect of irrigation on the survival of total coliforms in three semiarid soils after amendment with sewage sludge

Sewage sludges are increasingly used in soil amendment programmes, although not without risk since they contain, among other potential hazards, high concentrations of total coliform bacteria. In this paper we have studied the effect of irrigation on the survival of total coliforms in three semiarid degraded soils with different agricultural practices. Fresh sewage sludge was added at 50 g kg(-1) soil, and incubated in both the presence and absence of irrigation. The absence of irrigation led to a sharp decrease in the number of total coliforms in all soils, with the bacteria disappearing in 40 days. Irrigation produced a substantial initial increase in the number of coliforms in the three soils, although after 80 days there was none growing in any of the soils. The results showed that there were significant differences in the survival of coliform bacteria due to the presence or absence of irrigation.