Effect of fertilizers and irrigation on multi-configuration electromagnetic induction measurements

Electromagnetic induction (EMI) data are often used to investigate spatial and temporal patterns of soil texture, soil water content and soil salinity. We hypothesized that the EMI methodology might thus also offer potential to detect agricultural legacy effects originating from fertilizer application and irrigation of different fields. Therefore, we performed EMI measurements on two long-term field experiments (LTFE) at Thyrow near Berlin (Germany) that differed in agricultural management with regard to long-term irrigation in combination with mineral (NPK and lime) and organic amendments (straw and farmyard manure). Two different rigid-boom multi-coil EMI instruments were used to measure simultaneously the apparent electrical conductivity (ECa) over nine different depth ranges to study the entire soil profile from topsoil to deep subsoil. Additionally, soil samples were taken from the different treatments to ground-truth the measurements and disentangle the nutrient application or irrigation effects from natural soil heterogeneity. The soil samples indicated a rather homogenous soil and the correlation between soil parameters or states were not significant. However, the treatments showed significant differences in measured ECa values. In general, ECa values were largest on regularly irrigated as well as on mineral and organic fertilized plots, with regular irrigation exhibiting the largest impact on EMI records even though the last application was months before the EMI measurement. Overall, this study reveals that EMI data can support the classical in situ assessment of agricultural management effects within LTFE, while offering new potentials in detecting and understanding legacy effects of agricultural management on spatial soil properties at farm level.     

Differential alterations in the ability of bovine neutrophils to generate extracellular and intracellular reactive oxygen species during the periparturient period

The periparturient period of a dairy cow is associated with increased incidence and/or severity of certain infectious diseases, including mastitis. It is believed that the heightened physiological demands of calving and initiation of milk production contribute to a state of immunosuppression during this period. Previous studies have indicated that neutrophil production of reactive oxygen species (ROS), which is a critical element of the host innate immune response to bacterial infection, is impaired in the 1-2week period following calving. However, whether there is comprehensive inhibition of ROS production or selective inhibition of particular ROS remains unknown. The present study provides evidence that neutrophils isolated from cows (n=20) after calving have an increased capacity to generate intracellular ROS and an impaired ability to release extracellular superoxide anion and hydrogen peroxide.     

Partially defatted black soldier fly larva meal inclusion in piglet diets: effects on the growth performance,nutrient digestibility,blood profile,gut morphology and histological features

Background: The aim of this trial was to investigate the effects of different inclusion levels of a partial y defatted black soldier fly(BSF, Hermetia il ucens L.) larva meal on the growth performance, nutrient digestibility, blood profile, gut morphology and histological features of piglets. A total of 48 newly weaned piglets were individual y weighed(initial body weight(IBW): 6.1 ± 0.16 kg) and randomly al ocated to 3 dietary treatments(4 boxes as replicates/treatment and 4 animals/box). BSF larva meal was included at increasing levels(0% [BSF0], 5% [BSF5] and 10% [BSF10]) in isonitrogenous and isoenergetic diets formulated for two feeding phases: I(from d 1 to d 23) and I(from d 24 to d 61). The weight gain(WG), average daily gain(ADG), average daily feed intake(ADFI) and feed conversion ratio(FCR) were calculated for each feeding phase and for the whole trial. The haematochemical parameters and nutrient digestibility of the piglets were also evaluated. A total of 3 piglets per box were slaughtered on d 61 and the slaughtered piglets were submitted to morphometric investigations and histopathological examinations.Results: No overal significant differences were observed for growth performance(P 0.05), except for the ADFI of phase I, which showed a linear response to increasing BSF meal levels(P 0.05, maximum for the BSF10 group). Dietary BSF meal inclusion did not significantly influence the blood profile, except as far as monocytes and neutrophils are concerned,and these showed a linear and quadratic response, respectively, to increasing BSF meal levels(P 0.05, maximum for the BSF10 and BSF5 groups, respectively). On the other hand, the nutrient digestibility, gut morphology and histological features were not affected by dietary BSF meal inclusion(P 0.05).Conclusions: The obtained results show that a partial y defatted BSF larva meal can be used as a feed ingredient in diets for weaned piglets without negatively affecting their growth performance, nutrient digestibility, blood profile, gut morphology or histological features.     

Estimation of daily milk yield of Nellore cows grazing tropical pastures

Beef cows’ milk yield is typically determined by measuring milk yield once daily and then doubling this value to estimate daily production. However, it is not known whether this is accurate. Thus, we aimed to determine the association between morning and afternoon milk yield in grazing Nellore cows. Eighty Nellore cows were used, with initial weight of 516.0?±?1.0 kg. The experiment was a completely randomized factorial scheme, with 20 replications and four treatments (i.e., +?or ??pre-partum supplementation in combination with +?or ??post-partum supplementation): PRMM—1 kg of supplement/cow/day for 90 days pre-partum; MMPS—1 kg of supplement/cow/day for 90 days post-partum; PRPS—1 kg of supplement/cow/day for 90 days pre-partum and 90 days post-partum; and MM—only mineral mix ad libitum during pre- and post-partum. Milk was sampled on days 45, 135, and 225 post-partum (early, middle, and late lactation, respectively). No effects were observed of pre- and post-partum supplementation on milk yield (P?>?0.05). The afternoon/morning proportion of 0.45 in the early third of lactation was higher than other stages, which had a proportion of 0.41 (P?<?0.05). Post-partum supplementation increased milk protein in the morning and afternoon milking (P?<?0.05). There was also no effect of pre- and post-partum supplementation on afternoon-morning proportion other milk components (P?>?0.05). We conclude that estimating daily milk production of grazing beef cattle by multiplying a once daily milking amount times two is not accurate. Under the conditions of this study, proportion of total daily production represented by the ratio of afternoon/morning milking was 0.45 in early lactation (first third) and 0.41 in mid- and late lactation.     

Interaction of four monoterpenes contained in essential oils with model membranes: implications for their antibacterial activity

The present article reports the antimicrobial efficacy of four monoterpenes (thymol, carvacrol, p-cymene, and gamma-terpinene) against the Gram-positive bacterium Staphylococcus aureus and the Gram-negative bacterium Escherichia coli. For a better understanding of their mechanism of action, the damage caused by these four monoterpenes on biomembranes was evaluated by monitoring the release, following exposure to the compounds under study, of the water-soluble fluorescent marker carboxyfluorescein (CF) from large unilamellar vesicles (LUVs) with different lipidic composition (phosphatidylcholine, PC, phosphatidylcholine/phosphatidylserine, PC/PS, 9:1; phosphatidylcholine/stearylamine, PC/SA, 9:1). Furthermore, the interaction of these terpenes with dimyristoylphosphatidylcholine multilamellar vesicles as model membranes was monitored by means of differential scanning calorimetry (DSC) technique. Finally, the results were related also with the relative lipophilicity and water solubility of the compounds examined. We observed that thymol is considerably more toxic against S. aureus than the other three terpenes, while carvacrol and p-cymene are the most inhibitory against E. coli. Thymol and carvacrol, but not gamma-terpinene and p-cymene, caused a concentration-dependent CF leakage from all kinds of LUVs employed; in particular, thymol was more effective on PC and PC/SA LUVS than on PC/PS vesicles, while carvacrol challenge evoked a CF leakage from PC/PS LUVs similar to that induced from PC/SA LUVs, and lower than that measured with PC vesicles. Concerning DSC experiments, these four terpenes caused a decrease in Tm and (especially carvacrol and p-cymene) DeltaH values, very likely acting as substitutional impurities. Taken together, our findings lead us to speculate that the antimicrobial effect of thymol, carvacrol, p-cymene, and gamma-terpinene may result, partially at least, from a gross perturbation of the lipidic fraction of the plasmic membrane of the microorganism. In addition to being related to the physicochemical characteristics of the compounds (such as lipophilicity and water solubility), this effect seems to be dependent on the lipidic composition and net surface charge of the microbic membranes. Furthermore, the compounds might cross the cell membranes, thus penetrating into the interior of the cell and interacting with intracellular sites critical for antibacterial activity.     

Ion-trap tandem mass spectrometry for the analysis of polychlorinated dibenzo-p-dioxins, dibenzofurans, and dioxin-like polychlorinated biphenyls in food

This paper reports on the applicability of gas chromatography coupled to ion-trap tandem mass spectrometry (GC/ITMS/MS) for the analysis of polychlorinated dibenzo- p-dioxins (PCDDs), dibenzofurans (PCDFs), and dioxin-like polychlorinated biphenyls (dl-PCBs) in food. MS/MS parameters were selected to achieve the high sensitivity and selectivity required for food analysis. Good precision (RSD=5-18% for PCDD/Fs and 6-14% for dl-PCBs) and low limits of detection for PCDD/Fs (0.1-0.93 pg/g of fat) and dl-PCBs (0.1-0.89 pg/g of fat) were obtained. A comparative study of the congener-specific determination using both GC/ITMS/MS and GC-high resolution mass spectrometry (GC/HRMS) was performed by analyzing several matrices such as milk, fish oil, chicken, pork, fish, eggs, and a chicken compound feed, at low pg/g levels. The results using GC/ITMS/MS were in good agreement with those obtained by GC/HRMS. Consequently, GC/ITMS/MS is proposed for the analysis of PCDD/Fs and dl-PCBs in food and feed samples.     

Quantification of milk fat in chocolate fats by triacylglycerol analysis using gas-liquid chromatography

The development and in-house testing of a method for the quantification of milk fat in chocolate fats is described. A database consisting of the triacylglycerol profiles of 310 genuine milk fat samples from 21 European countries and 947 mixtures thereof with chocolate fats was created under a strict quality control scheme using 26 triacylglycerol reference standards for calibration purposes. Out of the individual triacylglycerol fractions obtained, 1-palmitoyl-2-stearoyl-3-butyroyl-glycerol (PSB) was selected as suitable marker compound for the determination of the proportion of milk fat in chocolate fats. By using PSB values from the standardized database, a calibration function using simple linear regression analysis was calculated to be used for future estimations of the milk fat content. A comparison with the widely used butyric acid method, which is currently used to determine the milk fat content in nonmilk fat mixtures, showed that both methods were equivalent in terms of accuracy. The advantage of the presented approach is that for further applications, i.e., determination of foreign fats in chocolate fats, just a single analysis is necessary, whereas for the same purpose, the C4 method requires two different analytical methods.     

Flavonoids from italian multifloral honeys reduce the extracellular ferricyanide in human red blood cells

In this study we investigated some biological properties of flavonoids recovered in the aqueous (AqE) and ether (EtE) extracts from four Italian multifloral honeys. In particular, a cell-free assay was employed to detect direct reduction of ferricyanide, whereas an assay using intact human erythrocytes was used to measure the ability to donate electrons to a trans-plasma membrane oxidoreductase. It was found that the AqE displays greater "in vitro" ferricyanide-reducing activity than the EtE but, unlike the latter, is virtually ineffective in the cell-based assay. Uptake studies employing high-performance liquid chromatography/mass spectrometry (HPLC/MS) showed that the different results were explained by the inability of AqE components to cross the erythrocyte plasma membrane and by the excellent uptake of EtE flavonoids, which, once within the cell, donate electrons to the membrane oxidoreductase to efficiently reduce extracellular oxidants. The latter property appears to depend on the content of ether-soluble flavonoids in the starting honeys.     

High-resolution protein structure determination by serial femtosecond crystallography

Structure determination of proteins and other macromolecules has historically required the growth of high-quality crystals sufficiently large to diffract x-rays efficiently while withstanding radiation damage. We applied serial femtosecond crystallography (SFX) using an x-ray free-electron laser (XFEL) to obtain high-resolution structural information from microcrystals (less than 1 micrometer by 1 micrometer by 3 micrometers) of the well-characterized model protein lysozyme. The agreement with synchrotron data demonstrates the immediate relevance of SFX for analyzing the structure of the large group of difficult-to-crystallize molecules.     

Qualitative and quantitative polymerase chain reaction (PCR) for detection of Leishmania in spleen samples from naturally infected dogs

Because infected dogs are widely considered to be the main domestic reservoir for Leishmania infantum (syn Leishmania chagasi) parasites in Brazil, the diagnosis of canine visceral leishmaniasis (CVL) must be made both accurately and promptly. The present study attempted to standardize a conventional polymerase chain reaction (cPCR) protocol for the detection of L. infantum DNA in canine spleen samples. Quantitative PCR (qPCR) technique was used to confirm the presence of Leishmania DNA in the canine spleen fragments. A comparison was made between the efficacies of these molecular diagnostic techniques and conventional parasitological and serological methods. cPCR protocols for spleen samples were standardized using primers that amplify a 145 bp fragment, located at the parasite kinetoplast minicircle. The genus specificity of the cPCR protocol was assessed by its inability to amplify the DNA of other common canine pathogens, such as Ehrlichia canis, Babesia canis, Toxoplasma gondii and Trypanosoma cruzi. cPCR protocol sensitivity was tested by assessing the reaction detection limit, determined to be 10 fg of L. infantum reference strain DNA, which corresponds to a range of 0.03-0.1 parasites per fragment. Standardized cPCR protocol was used to detect the presence of Leishmania in 45 dog spleen samples. Our results showed that 40% of the spleen fragment cultures were positive for Leishmania parasites, 58% of the dog serum samples tested positive using ELISA, and parasite DNA was detected in 44% using qPCR, while 47% of the spleen samples using cPCR. Diagnostic methods performance was assessed and revealed a better degree of ascertainment for cPCR when compared to other diagnostic methods. The sensitivity of ELISA was 83.3%, qPCR was 83.3%, and cPCR was 88.9%; PPV for ELISA was 57.7%, qPCR was 75% and cPCR was 76.2%; the Kappa coefficients were found to be 0.40 (fair) for ELISA, 0.64 (substantial) for qPCR and 0.68 (substantial) for cPCR. In both oligosymptomatic and polysymptomatic dogs, cPCR revealed the better performance analysis when compared to other diagnostic methods. The findings presented herein establish cPCR as the most indicated test to detect Leishmania when compared to the other two diagnostic methods evaluated. Despite the fact that the qPCR protocol provides a highly accurate quantification of parasites when targeting the SSU rRNA gene, this technique does not significantly improve the diagnosis of CVL when compared with the performance of the cPCR protocol, which focused on the kinetoplast minicircle.