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.     

Distribution of endocrine disruptors in the Mondego River estuary, Portugal

Recent studies in the Mondego River estuary demonstrated signs of pollution in the area, but the nature of the contamination remains unexplored. Because there seems to be any studies in that zone checking for the simultaneous presence of xenoestrogenic endocrine disrupting compounds, either of animal (estradiol and estrone), vegetal (daidzein, genistein and biochanin A), pharmaceutical (17alpha-ethynylestradiol) or industrial (bisphenol A, 4-octylphenol, 4-nonylphenol) origins, the main objective of this study was to investigate their presence, in every year season and at eight points along the estuarine gradient. For this propose, water samples (2 L) were collected in high and low tides, preconcentrated in the Oasis HLB cartridges and cleaned in silica cartridges before their analysis by HPLC-DAD and GC-MS. The current data showed the absence of contamination by animal or pharmaceutical estrogens and by alkylphenols, but demonstrated the presence of high levels of phytoestrogens (up to 1.1 mug/L) and of bisphenol A (up to 880.0 ng/L). Because these two chemicals existed in high environmental concentrations in the most upstream sampling station, and taking in account that these pollutants may additively contribute to endocrine disruption, namely on fish, it was concluded that at least this particularly area of the Mondego estuary deserves continuous monitoring programmes.     

Two-phase anaerobic digestion within a solid waste/wastewater integrated management system

A two-phase, wet anaerobic digestion process was tested at laboratory scale using mechanically pre-treated municipal solid waste (MSW) as the substrate. The proposed process scheme differs from others due to the integration of the MSW and wastewater treatment cycles, which makes it possible to avoid the recirculation of process effluent. The results obtained show that the supplying of facultative biomass, drawn from the wastewater aeration tank, to the solid waste acidogenic reactor allows an improvement of the performance of the first phase of the process which is positively reflected on the second one. The proposed process performed successfully, adopting mesophilic conditions and a relatively short hydraulic retention time in the methanogenic reactor, as well as high values of organic loading rate. Significant VS removal efficiency and biogas production were achieved. Moreover, the methanogenic reactor quickly reached optimal conditions for a stable methanogenic phase. Studies conducted elsewhere also confirm the feasibility of integrating the treatment of the organic fraction of MSW with that of wastewater.     

Towards Understanding the Impacts of the Pet Food Industry on World Fish and Seafood Supplies

The status of wild capture fisheries has induced many fisheries and conservation scientists to express concerns about the concept of using forage fish after reduction to fishmeal and fish oil, as feed for farmed animals, particularly in aquaculture. However, a very large quantity of forage fish is being also used untransformed (fresh or frozen) globally for other purposes, such as the pet food industry. So far, no attempts have been made to estimate this quantum, and have been omitted in previous fishmeal and fish oil exploitation surveys. On the basis of recently released data on the Australian importation of fresh or frozen fish for the canned cat food industry, here we show that the estimated amount of raw fishery products directly utilized by the cat food industry equates to 2.48 million metric tonnes per year. This estimate, plus the previously reported global fishmeal consumption for the production of dry pet food suggest that 13.5% of the total 39.0 million tonnes of wild caught forage fish is used for purposes other than human food production. This study attempts to bring forth information on the direct use of fresh or frozen forage fish in the pet food sector that appears to have received little attention to this date and that needs to be considered in the global debate on the ethical nature of current practices on the use of forage fish, a limited biological resource.     

Spatial configuration of land-use/land-cover in the Pujal-Coy project area, Huasteca Potosina region, Mexico

This article analyzes the relationship between the configuration and spatial reorganization of land-use and land-cover in the Pujal-Coy project area, Huasteca Potosina region, eastern San Luis Potosí, Mexico, as well as the relationship between these changes and the environmental conditions prevailing in the area. Land-use and land-cover changes were determined through the analysis and interpretation of satellite images from different dates. The changes identified in the different study periods were correlated with the prevailing physical factors. The results show that the spatial configuration of farming activities, initially induced by the implementation of a regional development project, is highly correlated to the presence of limiting factors such as soil type, slope, and climate. Particularly, the former represents the element that has led to the establishment of the current distribution pattern of farming activities.     

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.     

Profiling the spatial concentration of allethrin and piperonyl butoxide using passive sorptive sampling and thermal desorption capillary GC-MS

Spatial concentration distribution of a chemical in an indoor environment is an important factor in the evaluation of chemical nuisances. However, straightforward techniques for the determination of this distribution are not very common and usually limited in their application. Sorptive sampling using polydimethylsiloxane-coated stir bars and the combination of active and diffusive sampling were shown to allow uncomplicated spatial concentration profiling of multiple compounds in an indoor environment. The validity of the approach was demonstrated in the analysis of the spatial concentration distribution of a pyrethroid insecticide in a common bedroom. The relative concentrations of allethrin and piperonyl butoxide were profiled throughout the room upon the application of an insecticide vaporizer.     

Maternal transfer of organochlorine compounds to oocytes in wild and farmed gilthead sea bream (Sparus aurata)

To investigate the transfer of lipophilic organochlorine compounds to the gonads in gilthead sea bream females, liver and oocytes from wild and farmed females, just before spawning, were analysed by gas chromatography coupled to tandem mass spectrometry after clean-up of the fatty extracts by normal phase HPLC. This analytical methodology allowed us to achieve limits of detection of around 0.1 ng/g. Data obtained show that the chemicals studied are transferred from the liver to oocytes in ratios up to 0.51. In general this means that less than a half of pollutant load is transferred from liver to oocytes. A significant correlation (p<0.05) between log K(ow) and ratios (oocytes/liver) was found. Compounds with larger Log K(ow) present lower lipid-based concentration ratios (oocytes/liver). We have inferred that the reason the fugacity model is not accomplished is specifically dependent on the vitellogenesis process, which involves the transfer of lipids from the liver to oocytes in teleostei. No significant differences (t-Student) have been found between wild and farmed females transfer ratios (oocytes/liver).     

Empirical relationship between precision and ultra-trace concentrations of PCDD/Fs and dioxin-like PCBs in biological matrices

Dioxin analysis in food and feed can be characterized as an analytical application where very high accuracy is required at very low levels of contamination. Gas chromatography (GC) in combination with 13C-label isotope dilution (ID) high resolution mass spectrometry (HRMS) is the reference congener-specific technique characterized by pronounced selectivity, precision and trueness at parts-per-trillion (ppt) and sub-parts-per-trillion (sub-ppt) levels. The quality of the analytical data produced routinely by a laboratory should be adequate for its intended purpose, i.e., one will seek a compromise between the cost and time needed and the consequences of incorrect decisions due to erroneous results. The requirements for reproducibility are usually dependent on the analyte concentrations and have been expressed in various empirical functions. While Horwitz or modified functions are widely useful for many purposes, it would be difficult to expect these functions to cover every analytical problem. This study reports on precision characteristics achieved by the GC-ID-HRMS reference method for polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and dioxin-like polychlorinated biphenyls (DL-PCBs) in food and feed in two interlaboratory method-performance studies among expert laboratories with long-standing experience in this field. Striking linear functions in log scale between reproducibility standard deviation and congener's level over a concentration range of 10(-8)-10(-14)g per g fresh weight are observed. The data fit very well to a Horwitz-type function of the form sR=0.153c0.904, where sR and c are dimensionless mass ratios expressed in pg g(-1) on fresh weight, regardless of the nature of the toxic congeners, food and feed matrices, or sample preparation methods. We demonstrate that the proposed function is suitable for use as a fitness-for-purpose criterion for proficiency assessment in interlaboratory comparisons on dioxins and related compounds in food.