Characterization of fluoroquinolone resistance in Escherichia coli strains from ruminants.

Thirty-seven fluoroquinolone-resistant Escherichia coli strains from ruminants (according to Clinical and Laboratory Standards Institute guidelines) were screened by molecular methods for mutations in the quinolone resistance-determining region (QRDR) of the gyrA and parC genes and for the presence of the qnrA gene. One of the strains studied was an enterohemorrhagic E. coli (EHEC) strain potentially pathogenic for humans. Three E. coli strains resistant to enrofloxacin (minimal inhibitory concentration [MIC] = 2 microg/ml) but not to ciprofloxacin (MIC = 1 microg/ml) presented single mutations in the gyrA and parC genes, while 34 strains resistant to both fluoroquinolones presented double and single mutations in gyrA and parC, respectively (31 strains), or double mutations in gyrA and parC (3 strains). The EHEC strain presented a double amino acid substitution in the GyrA protein (Ser-83-->Leu and Asp-87-->Gly) and a double amino acid substitution in the ParC protein (Gly-78-->Cys and Ser-80-->Arg), one of which has not been previously described. The present study shows that most of the mutations in the QRDR of the gyrA and parC genes of fluoroquinolone-resistant E. coli strains from ruminants are the same as those seen in E. coli strains from other animal species and humans and that there are no differences in mutation patterns in the QRDR of E. coli strains from healthy ruminants and those with diarrhea. No strains carried qnrA, which indicates that this gene does not play an important role in the selection of fluoroquinolone-resistant E. coli strains from ruminants.     

Nursery areas of Micromesistius poutassou and Sardina pilchardus unveil their reproductive strategies in the northwestern Mediterranean Sea

Winter conditions in the NW Mediterranean cause instability of the water column and non-geostrophic dynamics, such as vertical mixing and convection are significant. These events involve nutrient supply to the photic zone that can sustain high productivity. In this study, we aim to investigate the role of winter hydrodynamics on the spawning strategies of Sardina pilchardus and Micromesistius poutassou. Data were obtained on two oceanographic cruises (February 2017 and 2018) off the Catalan coast. The occurrence of S. pilchardus eggs very close to the coast indicated a clear preference of the species for spawning in coastal areas. Preflexion and postflexion larvae exhibited a slightly wider distribution showing a clear association with the cold, less saline and more productive coastal waters. Preflexion larvae of M. poutassou were found on the upper slope and over the shelf, being offshore limited by the shelf/slope front present all along the slope. The front would act as a barrier preventing their dispersion towards the open sea. M. poutassou larvae in advanced developmental stages were located close to the coast in the productive shelf waters, with instabilities of the front contributing to larval transport from offshore waters to the coast. The vertical distribution of both species showed high variability, not only related with the daily cycle or developmental stage, but also with the vertical structure of the water column. Overall, the results provide some clues on how the spawning strategies of both species may evolve under future scenarios of higher winter-stratification, because of the global warming.     

Selective effects of forest fires on the structural domains of soil humic acids as shown by dipolar dephasing <Superscript>13</Superscript>C NMR and graphical-statistical analysis of pyrolysis compounds

Purpose

Data management strategies of pyrolysis results and NMR acquisition modes were examined in humic acids (HAs) from control soils and fire-affected soils. The information supplied by dipolar dephasing (DD) ¹³C NMR spectroscopy and Curie-point pyrolysis were used to assess chemical structures hardly recognizable and measurable, or of unclear interpretation, when using ¹³C NMR under standard acquisition pulses (cross-polarization/magic angle spinning, CPMAS).

Materials and methods

The HAs were isolated from two forest soils under Pinus halepensis and Pinus sylvestris in control and burned sites affected by medium or severe-intensity wildfires. For NMR analyses, during DD acquisition conditions, a 180° ¹³C pulse was inserted to minimize phase shifts. Curie-Point pyrolysis was carried out at 510 °C for 5 s, and the pyrolysis fragments were analyzed by GC/MS. The total abundances of the major pyrolysis products were compared by an update of the classical Van Krevelen’s graphical-statistical approach, i.e., as surface density values in the space defined by the compound-specific H/C and O/C atomic ratios.

Results and discussion

The DD ¹³C NMR experiments displayed significant differences in the HA spectral profiles as regards to the standard CPMAS ¹³C NMR acquisition conditions, mainly in the chemical shift region of alkyl structures as well as for tannin- or carbohydrate-like O-alkyl structures. In fact, the comparison between DD and CPMAS solid-state NMR suggested shortening of alkyl chains and generation of carbohydrate-derived, unsaturated structures—viz. furans—which adds to the aromatic domain. Pyrolytic results showed fire-induced specific changes in HAs chemical structure and its molecular diversity. The changes were evident in the location and sizes of the different clusters of pyrolysis compounds defined by their atomic ratios.

Conclusions

The DD ¹³C NMR provided specific information on the fate of aliphatic structures and the origin of unsaturated HA structures, which could be helpful in differentiating “inherited” from “pyrogenic” aromatic structures. This is further confirmed by the analysis of the molecular assemblages of pyrolytic products, which showed accumulation of condensed polyaromatic domains in the HAs after the high-intensity fire, accompanied by a recalcitrant alkyl hydrocarbon domain. Medium-intensity fire led to aromaticity increase due to a selective accumulation of lignin-derived phenols concomitant to the depletion of aliphatic hydrocarbon constituents.

     

Competitive ability of ALS‐inhibitor herbicide‐resistant Fimbristylis miliacea

Fimbristylis miliacea, a weed in rice, has evolved resistance to acetolactate synthase (ALS) inhibitors. This study aimed to investigate the competitive abilities of ALS‐resistant (R) and ALS‐susceptible (S) F. miliacea with rice. A replacement series experiment was conducted in the glasshouse at the Federal University of Pelotas, Brazil. The proportions of rice to F. miliacea were 100:0, 75:25, 50:50, 25:75 and 0:100, with 1060 plants m^?2. The experimental units were arranged in a completely randomised design with four replications. A follow‐up study was conducted at the University of Arkansas, Fayetteville, USA, in a split‐plot design with four replications. The main plot was species mixture (rice × R, rice × S, R × S). The subplot was competition partitioning (below‐ and above‐ground, below‐ground only, above‐ground only and no interspecific competition). Leaf area, plant height and shoot dry mass were recorded. Rice was more competitive than the R or S F. miliacea. In equal proportions of rice and F. miliacea, regardless of ecotype, the relative leaf area, height and dry mass of rice were greater than that of F. miliacea. The ALS‐resistant ecotype was less competitive with rice than the S ecotype. Intraspecific competition among rice plants was stronger than rice competition with F. miliacea. Competition for below‐ground resources was the most critical aspect of interference among rice and F. miliacea. In production fields, high infestation levels of F. miliacea results in significant yield losses; thus, resistance to ALS inhibitors needs to be curtailed.     

Determination of volatile organic compound patterns characteristic of five unifloral honey by solid-phase microextraction-gas chromatography-mass spectrometry coupled to chemometrics

We report the evaluation of the floral origin of honey by analysis of its volatile organic compounds (VOCs) profile, joined with the use of combined pattern recognition techniques. Honey samples, from five floral origins, were analyzed by headspace solid-phase microextraction-gas chromatography-mass spectrometry, selecting 35 VOCs out of the entire profiles, which were analyzed by hierarchical cluster analysis (HCA), stepwise discriminant analysis (SDA), and K-nearest-neighbor (KNN). Both HCA and SDA were used as exploratory tools to select a group of VOCs representing similitude and differences among studied origins. Thus, six out of 35 VOCs were selected, verifying their discriminating power by KNN, which afforded 93% correct classification. Therefore, we drastically reduced the amount of compounds under consideration but kept a good differentiation between floral origins. Selected compounds were identified as octanal, benzeneacetaldehyde, 1-octanol, 2-methoxyphenol, nonanal, and 2-H-1-benzopyran-2-one. The analysis of VOC profiles, coupled to HCA, SDA, and KNN, provides a feasible alternative to evaluate the botanical source of honey.     

Efficacy of five selected acaricides against Tetranychus urticae (Acari: Tetranychidae) and their side effects on relevant natural enemies occurring in citrus orchards

BACKGROUND: Three groups of natural enemies are fundamental in citrus IPM in Spain: coccinellid and phytoseiid predators and hymenopteran parasitoids. Tetranychus urticae Koch is an important pest affecting citrus, for which biological control has not yet been achieved; therefore, acaricides are commonly used to control it. The goal of this study was to measure the efficacy of different acaricides on this mite and their side effects on three natural enemies relevant for citrus (Cryptolaemus montrouzieri Mulsant, Neoseiulus californicus McGregor and Aphidius colemani Viereck).RESULTS: Some products proved highly effective against T. urticae and harmless to A. colemani (mineral oil, tebufenpyrad, clofentezine and fenazaquin). However, almost all products tested were slightly harmful for both the predators considered. Fenazaquin was even moderately harmful for N. californicus. CONCLUSION: Further studies, like that presented here, are necessary to gain a better understanding of integrating biological and chemical controls. When considering both efficacy and side effects on beneficial arthropods, the best options would seem to be mineral oil, tebufenpyrad and clofentezine. However, it is urgent to complete testing of the side effects of the acaricides used in citrus. This question is crucial if the fact that two recently introduced Tetranychidae are being controlled in citrus by chemical means exclusively is considered.