Detection of Listeria monocytogenes in ready-to-eat food by Step One real-time polymerase chain reaction

The aim of this study was to follow contamination of ready-to-eat food with Listeria monocytogenes by using the Step One real time polymerase chain reaction (PCR). We used the PrepSEQ Rapid Spin Sample Preparation Kit for isolation of DNA and MicroSEQ? Listeria monocytogenes Detection Kit for the real-time PCR performance. In 30 samples of ready-to-eat milk and meat products without incubation we detected strains of Listeria monocytogenes in five samples (swabs). Internal positive control (IPC) was positive in all samples. Our results indicated that the real-time PCR assay developed in this study could sensitively detect Listeria monocytogenes in ready-to-eat food without incubation.     

Temporal and spatial changes of PAH concentrations in Mytilus galloprovincialis from Ria de Vigo (NW Spain)

The aim of this study was to establish the temporal trends and spatial distribution of Polycyclic Aromatic Hydrocarbons (PAHs) in Ria de Vigo by means of studying the PAH burden in wild mussels. The samples were collected in eight sites along the margins of the Ria de Vigo during November from 1998 to 2008 and Polycyclic Aromatic Hydrocarbon contents were determined using high performance liquid chromatography–fluorescence derivatization (HPLC–FLD). Quality of chemical analysis was guaranteed by participation in QUASIMEME intercalibration exercises carried out during the period of analysis. The concentrations for the sum of 13 PAHs were in the range 24–480 μg/kg dw, typical of an urban and industrialized area (50–500 μg/kg dw) except for a punctual input in La Guía in 1998 that led to a concentration above 1,000 μg/kg dw. In general, the sites in the outermost section of the estuary (ría) showed the lowest values, with an increasing gradient in the medium and inner parts of the ría. Temporal patterns show, in general, a downward trend. The relative proportion of 3 and 4 ring PAHs also varies with the site, the former being predominant in the outermost area and the latter in the medium and inner parts of the ría. This is consistently related with the sources and processes taking place in different areas of the estuary.     

Assessment of oxidative stress and metabolic changes in common carp (Cyprinus carpio) acutely exposed to different concentrations of the fungicide tebuconazole

The aim of this research is to determine the lethal concentration (LC(50-96 h)) of fungicide tebuconazole for common carp (Cyprinus carpio) and to investigate the occurrence of oxidative stress and metabolic changes after acute exposure to different concentrations of tebuconazole. For this purpose, several parameters were assessed as whole-body levels of thiobarbituric acid-reactive substance (TBARS), protein carbonyl, catalase, superoxide dismutase, glutathione S-transferase, nonprotein thiols, ascorbic acid, glycogen, glucose, lactate, protein, amino acids and ammonia in tebuconazole-exposed fish. The calculated LC(50-96 h) was 2.37 mg L(-1). Fish exhibited significant increase in TBARS levels in all concentrations used while the enzymatic and nonenzymatic antioxidants were decreased. Among the metabolic parameters, glycogen and glucose increased at 1.19 mg L(-1) and protein levels decreased at 1.78 and 2.37 mg L(-1). In conclusion, the fish health was adversely affected by exposure to tebuconazole, and those changes can compromise animal survival in the natural environment. The results indicate that the some of the parameters measured like a possible biomarkers of exposure to tebuconazole for this species of fish.     

Environmental release and mass flux partitioning of PCDD/Fs during normal and transient operation of full scale waste to energy plants

The paper reports on global release and mass partitioning in the flux of residues of PCDD/Fs, evaluated with dedicated field campaigns at a municipal solid waste incineration plant during normal and transient operation. Results are compared with those obtained in other installations equipped with furnaces, energy recovery options and flue gas treatment technologies representative of most of the European incineration plants currently in operation. Levels of the pollutants of interest were determined in all the solid, liquid and gaseous residues produced by every single facility, and the results analysed in terms of the effects arising from the fed waste and the configuration of the plant. PCDD/Fs total release between 1.5 and 45 microg I-TEQ per ton of burned waste was evaluated, with lower values resulting from the adoption of catalytic conversion process for flue gas treatment. Most of the mass flux emitted is associated with solid residues deriving from activated carbon PCCD/F dry removal options, with significant contributions also from fly ash produced by particulate removal devices located immediately downstream the boiler and from scrubber blowdowns treatment sludge. During transient operating conditions the dioxin total release may increase by 50% with comparison to steady-state functioning.     

Stoichiometric controls of nitrogen and phosphorus cycling in decomposing beech leaf litter

Resource stoichiometry (C:N:P) is an important determinant of litter decomposition. However, the effect of elemental stoichiometry on the gross rates of microbial N and P cycling processes during litter decomposition is unknown. In a mesocosm experiment, beech (Fagus sylvatica L.) litter with natural differences in elemental stoichiometry (C:N:P) was incubated under constant environmental conditions. After three and six months, we measured various aspects of nitrogen and phosphorus cycling. We found that gross protein depolymerization, N mineralization (ammonification), and nitrification rates were negatively related to litter C:N. Rates of P mineralization were negatively correlated with litter C:P. The negative correlations with litter C:N were stronger for inorganic N cycling processes than for gross protein depolymerization, indicating that the effect of resource stoichiometry on intracellular processes was stronger than on processes catalyzed by extracellular enzymes. Consistent with this, extracellular protein depolymerization was mainly limited by substrate availability and less so by the amount of protease. Strong positive correlations between the interconnected N and P pools and the respective production and consumption processes pointed to feed-forward control of microbial litter N and P cycling. A negative relationship between litter C:N and phosphatase activity (and between litter C:P and protease activity) demonstrated that microbes tended to allocate carbon and nutrients in ample supply into the production of extracellular enzymes to mine for the nutrient that is more limiting. Overall, the study demonstrated a strong effect of litter stoichiometry (C:N:P) on gross processes of microbial N and P cycling in decomposing litter; mineralization of N and P were tightly coupled to assist in maintaining cellular homeostasis of litter microbial communities.     

The use of polymerase chain reaction for the identification of sciaenid eggs

Traditionally, sciaenid eggs have been identified based on morphological characteristics such as size, number of oil globules and/or pigmentation patterns. Identification of sciaenid eggs by these procedures is time consuming and often inaccurate due to considerable egg size overlap among species. The utilization of molecular techniques for the identification of economically important species has become a fundamental component in ecological studies involving fish eggs and larvae. This study reports the development of a series of both multiplex and individual polymerase chain reactions to identify the eggs of 11 sciaenid species commonly found in the Gulf of Mexico and estuaries near Port Aransas and Corpus Christi, TX, USA. Following method development, the discriminatory power of the assay was first determined with samples from adult fish collected from Aransas and Corpus Christi Bays, Galveston Bay and the lower Laguna Madre in northern Mexico. Most (97%) of these fishes were correctly identified to the level of species. To demonstrate the applicability of the assay, wild fish eggs were collected and analyzed from the Aransas Pass tidal inlet from September through December 2005. During this period, the eggs of four target species were positively identified which was in keeping with current knowledge regarding the spawning areas and seasons of these sciaenids based on the presence of mature females, eggs and/or larvae. Future use of this method can provide valuable information to better discriminate spawning sites and seasons of these species.     

Terbuthylazine persistence in an organic amended soil

The aim of this work was to study the effect of the application of a solid waste from olive oil production (alperujo) on the movement and persistence of the herbicide terbuthylazine (N2-tert-butyl-6-chloro-N4-ethyl-1,3,5-triazine-2,4-diamine). An experimental olive grove was divided in two plots: (i) Plot without organic amendment (blank) and (ii) Plot treated with alperujo during 3 years at a rate of 17920 kg of alperujo ha(- 1). Terbuthylazine was applied to both plots at a rate of 2 kg ha(- 1) a.i. Triplicates from each plot were sampled at 3 depths (0-10, 10-20 and 20-30 cm), air-dried, remains of olive leaves, grass roots, and stones removed and sieved through a 5 mm mesh sieve. Terbuthylazine was extracted with methanol 1:2 weight:volume ratio, the extracts were evaporated to dryness, resuspended in 2 mL of methanol, filtered and anylized by high performance liquid chromatography (HPLC). Higher amounts of terbuthylazine were detected at each sampling depth in plots treated with alperujo. The increase in soil organic matter content upon amendment with alperujo slightly increased sorption, suggesting that other factors beside sorption affect terbuthylazine degradation rate in organic amended soils.     

Soil carbon dynamics in a Mediterranean forest during the Kyoto Protocol commitment periods

The purpose of the present work is to asses the possibility of detecting changes in soil organic carbon (SOC) at the end of the 5-years of the first Commitment Period (CP) of the Kyoto Protocol of the United Nation’s Framework Convention on Climate Change (1 January, 2008–31 December, 2012), by both direct measurement and the use of an opportunely evaluated SOC model, CENTURY. The investigated soil is young, developed since 28 years on virtually C-free spoil banks and under the influence of two managed forest stands, one a mix of English oak (Quercus robur L.) and Italian alder (Alnus cordata Loisel.) and the other pure English oak. The SOC stock of either stand was monitored since the time the stands were planted in 1981, and it was used together with other parameters for the model evaluation, while the future projections for the end of the first (2012) and second (2017) CP were made according to two extreme IPCC climatic scenarios: A1F1, the most dramatic, and B2, among the less impacting. Direct SOC measurements performed at the beginning and at the end of a time frame equivalent to a commitment period (2004–2008) had not shown significant variations in either stands. Compared to the 2008 SOC stock, in both stands the model shows variations at the end of the first CP from 0.7 to 1.8 Mg C ha⁻¹ for the A1F1 scenario and from 0.3 to 1.7 Mg C ha⁻¹ for the B2. These variations are within the standard deviations of the C stocks measured in 2008. On the contrary, at the end of the second CP, the modelled SOC increments range from 2.5 to 3.6 Mg C ha⁻¹ (A1F1) or from 1.9 to 3.4 Mg C ha⁻¹ (B2), indicating the possibility to detect the SOC changes by direct measurement, since the values well agree with the minimum detectable variation estimated for both sites in 3.3–4.5 Mg C ha⁻¹. This work shows that SOC stock changes measured directly in the field can be minimal at the end of both CPs, and that CENTURY well simulates the SOC dynamics of the stands. The use of such a model, validated at long-term experimental sites, hence represents an effective tool for estimating future changes in SOC amounts in support of direct measurements when a short period of time, such as the CP, is considered.     

Investigation of ZnO nanoparticles’ ecotoxicological effects towards different soil organisms

Introduction  

Nanomaterials have widespread applications in several industrial sectors. ZnO nanoparticles (NPs) are among the most commonly used metal oxide NPs in personal care products, coating and paints. However, their potential toxicological impact on the environment is largely unexplored.     

Monitoring of PCDD/Fs and dioxin-like PCBs and seasonal variations in mussels from the Mar Grande and the Mar Piccolo of Taranto (Ionian Sea,Southern Italy)

The levels and specific profiles of polychlorinated dibenzo-p-dioxins, dibenzofurans (PCDD/Fs) and dioxin-like polychlorinated biphenyls (DL-PCBs) in Mytilus galloprovincialis from the Mar Grande and the Mar Piccolo of Taranto were determined during the extensive monitoring plan of Local Health Authority to assess PCDD/Fs and DL-PCBs contamination in food and farm products, within 20 km from the industrial areas of Taranto, between March and December 2011. The average Total Toxicity Equivalence (TEQ) values for the sum of PCDD/F and DL-PCBs ranged from 1.61 to 5.63 pg WHO₂₀₀₅-TEQ g^?1 wet weight basis, with the highest in the first inlet of the Mar Piccolo. In particular, DL-PCBs were the dominant chemicals in all samples, followed by PCDFs and PCDDs. Congener patterns in mussels were similar, indicating a homogeneous behavior in studied areas and, probably, the same type of source. The seasonal concentrations trend showed a relevant increase of dioxin and dioxin-like PCBs TEQs during the summer months, exceeding the limits set by the European Community for food and foodstuff. Reducing PCDD/Fs and PCBs is necessary to decrease contamination levels in order to safeguard marine ecosystem and human health in the Taranto area.