SYBR green real time-polymerase chain reaction as a rapid and alternative assay for the efficient identification of all existing Escherichia coli biotypes approved directly in wastewater samples

Escherichia coli has been recognized as the principal indicator of fecal contamination of water. Indeed, E. coli is the only species in the coliform group found in relationship with gastrointestinal tract of human and warm‐blooded animals and subsequently excreted in large numbers in the human feces. To obtain a complete picture of water quality and therefore, a better protection of public health, different techniques for water analysis have been proposed. In this article, we describe an alternative method that uses SYBR green real time‐polymerase chain reaction (RT‐PCR) technology to identify and quantify all E. coli biotypes in a group of wastewater samples collected from a wastewater depurator located in South of Italy. This new RT‐PCR protocol is accurate in measuring the concentration of chromosomal E. coli DNA using the amplification of three new specific fragments of the following bacteria genes: CadC, HNS, and Allan whose sequence is specific for E. coli family and conserved in all E. coli subtypes. This method allowed us to detect the presence of all E. coli biotypes directly in wastewater samples and estimated the correspondence between colony forming units and bacterial DNA concentrations. The availability of a rapid and sensitive method may be useful to monitor the persistence of E. coli in water, to evaluate the efficiency of wastewater purification treatments and the possible recycle for agricultural use. Furthermore, the development of a simple and routine method to monitor water quality with RT‐PCR analysis can encourage the testing of a higher number of samples. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 28: 1106–1113, 2012     

Enhancement of Glutaraldehyde Biocidal Efficacy by the Application of an Electric Field. Effect on Sessile Cells and on Cells Released by the Biofilm

Summary The aim of this paper was to evaluate the possible enhancement of the biocidal efficacy of glutaraldehyde against Pseudomonas fluorescens biofilms by the application of an electric field. The behaviour of sessile cells and cells released by the biofilms was assed. Biofilms were formed on thin stainless steel coupons immersed in culture media inoculated with Pseudomonas fluorescens. Treatments using glutaraldehyde (TGA) and both glutaraldehyde and electric field application (TGAEF) were carried out with the samples with biofilms. TGA: samples with biofilms were immersed in glass cells containing a buffer solution with different glutaraldehyde concentrations in the 25–500 ppm range. TGAEF: samples with biofilms were immersed in an electrochemical cell containing glutaraldehyde solution where a direct electric current (4 × 10⁻⁴ A cm⁻²) was delivered to the chamber. The evolution of biofilms was observed through optical microscopy at real time. Results show that the electric field enhanced glutaraldehyde efficacy reducing the number of surviving cells in the range of one to four orders with respect to those with TGA treatment. The sensitivity of the cells to the treatments decreased in the following order: planktonic cells > cells released by the biofilm > sessile cells.     

Fatal Pulmonary Mucormycosis due to <Emphasis Type="Italic">Rhizopus homothallicus</Emphasis>

We report here a case of cavitary pneumonia due to Rhizopus homothallicus in a diabetic patient. This is the first proven case of R. homothallicus infection in Western countries and the third case described worldwide. The organism was isolated from lung biopsy and identified after amplification and sequencing of the internal transcribed spacer region.     

Flower scent bouquet variation and bee pollinator visits in <Emphasis Type="Italic">Stevia rebaudiana</Emphasis> Bertoni (Asteraceae), a source of natural sweeteners

Pollinators provide a key service to both natural and agricultural ecosystems. Little is reported on the pollination chemoecology of Stevia rebaudiana (Asteraceae), a hermaphroditic species producing self-incompatible florets in small corymbs. We investigated the chemistry of volatiles potentially involved in its pollination system. The VOCs emitted by the corymbs of 27 F1 open-pollinated genotypes were collected by solid-phase micro-extraction and analyzed by gas chromatography–mass spectrometry (GC–MS), as well as morphometric data of the genotypes were recorded. Finally, we quantified the abundance of pollinators for each genotype. S. rebaudiana flowers were mainly visited by bees (Apidae and Halictidae), followed by hoverflies (Diptera: Syrphidae). GC–MS indicated that S. rebaudiana was characterized by a complex scent profile with large variability among F1 plants. Discriminant analysis showed that limonene, δ-elemene and bicyclogermacrene were the compounds explaining most of the scent bouquet difference between high attractive (>40 pollinators/plant) from low attractive pollinator power (<40 pollinators/plant). Limonene was the most representative VOC among plants that are more attractive to pollinators, while high emissions of δ-elemene and bicyclogermacrene were linked to plants that are less attractive to pollinators. S. rebaudiana morphometric data highlighted that, besides floral VOCs, corymb abundance and size, as well as plant height, may route pollinator visits. Overall, this study adds knowledge on floral phenology and pollinator ecological traits of S. rebaudiana, allowing a deeper understanding of its chemical ecology and pollination.     

Downregulation of GABA<Subscript>A</Subscript> Receptor Recycling Mediated by HAP1 Contributes to Neuronal Death in In Vitro Brain Ischemia

Downregulation of GABAergic synaptic transmission contributes to the increase in overall excitatory activity in the ischemic brain. A reduction of GABA_A receptor (GABA_AR) surface expression partly accounts for this decrease in inhibitory activity, but the mechanisms involved are not fully elucidated. In this work, we investigated the alterations in GABA_AR trafficking in cultured rat hippocampal neurons subjected to oxygen/glucose deprivation (OGD), an in vitro model of global brain ischemia, and their impact in neuronal death. The traffic of GABA_AR was evaluated after transfection of hippocampal neurons with myc-tagged GABA_AR β3 subunits. OGD decreased the rate of GABA_AR β3 subunit recycling and reduced the interaction of the receptors with HAP1, a protein involved in the recycling of the receptors. Furthermore, OGD induced a calpain-mediated cleavage of HAP1. Transfection of hippocampal neurons with HAP1A or HAP1B isoforms reduced the OGD-induced decrease in surface expression of GABA_AR β3 subunits, and HAP1A maintained the rate of receptor recycling. Furthermore, transfection of hippocampal neurons with HAP1 significantly decreased OGD-induced cell death. These results show a key role for HAP1 protein in the downmodulation of GABAergic neurotransmission during cerebral ischemia, which contributes to neuronal demise.     

A comparative life cycle assessment of the sugarcane value chain in the province of Tucumán (Argentina) considering different technology levels

Purpose

The purpose of this work is to quantify the environmental impact of the sugarcane industry in Tucumán (Argentina) through the life cycle analysis (LCA). The distinctive feature is the consideration of different technology levels (TLs) in the agricultural stage: high (HTL), medium (MTL), and low (LTL).

Methods

The scope of the study covers the agricultural and industrial stages through a “from cradle to gate” approach (from sugarcane cultivation until production of finished products: sugar and alcohol). The system is divided into Agriculture, Sugar Factory, and Distillery. Data used for the inventory are mainly provided by local experts, sugarcane growers, and processing companies. The characteristics of each TL are taken from a regional classification. For the impact assessment, the CML 2001 model (nine impact categories) is used.

Results and discussion

Regardless of the TL, in most of the impact categories, an important contribution attributable to the use of synthetic agrochemicals is evident. As for the comparison among TLs, the ethanol produced with HTL has less impact values than the ones produced with MTL and LTL in seven categories. These results can be mainly explained by the better cultural yields obtained with HTL, and to the fact that sugarcane is not burnt before harvesting in HTL as it is in MTL and LTL.

Conclusions

This study explores the implications of using different TLs for the agricultural tasks on the sugarcane supply chain in Tucumán, which is characterized by a vertically nonintegrated productive scheme. If practices associated to HTL are implemented, a reduction of the environmental impact is observed in most categories. It is necessary to compare these results with economic and social implications to ensure sustainability of the sugarcane value chain.

     

Effects of strength training with elastic band programme on fitness components in young female handball players: a randomized controlled trial

This study examined the effect of a 10-week programme of strength training with elastic band (STEB) on fitness components in young female handball players. Twenty-six young female handball players (aged 15.8 ± 0.2 years) from the same club participated in this study. They were randomly assigned between experimental (EG; n = 13) and control (CG; n = 13) groups. The EG performed the STEB, replacing some handball-specific drills in the regular handball training. The CG followed the regular handball training (i.e., mainly technical-tactical drills, small sided and simulated games, and injury prevention drills). Two-way analyses of variance were used to assess: handgrip; back extensor strength; medicine ball throw; 30 m sprint times; Modified Illinois change-of-direction (Illinois-MT); four jump tests: squat jump (SJ), countermovement jump (CMJ), countermovement jump with arm swing (CMJA) and five-jump test (5JT); static (Stork test) and dynamic balance (Y Balance Test); and repeated sprint T-test (RSTT). Results revealed significant gains in handgrip - right (p < 0.001, d = 1.75: large), handgrip - left (p < 0.001, d = 2.52: large), back extensor (p < 0.001, d = 2.01: large), and medicine ball throw (p = 0.002, d = 0.95: large) with EG compared to the CG. The EG also demonstrated greater improvement in sprint performance over 20 m (Δ = 10.6%, p = 0.001, d = 1.07: large) and 30 m (Δ = 7.2%, p < 0.0001, d = 1.56: large) compared to the CG. The EG showed better Illinois-MT (Δ = 5.6%, p = 0.034, d = 0.62: medium) compared to the CG. Further, EG posted significant improvements in the SJ (Δ = 17.3%, p = 0.048, d = 0.58: medium), CMJ (Δ = 17.7%, p = 0.017 d = 0.71: medium), and CMJA (Δ = 16.3%, p = 0.019, d = 0.69: medium) compared to the CG. Similarly, the EG exhibited significant improvement in RSTT best time [p = 0.025, d = 0.66 (medium)], RSTT mean time [p = 0.019, d = 0.69 (medium)] and RSTT total time [p = 0.019, d = 0.69 (medium)] compared to the CG. In conclusion, the 10-week STEB improved the physical abilities in young female handball players.