Slow nitrogen release from humic substances modified with aminoorganosilanes

Purpose

The purpose of the present study is to evaluate slow-release nitrogen capabilities of soil amendments obtained by modification of humic materials from peat and lignite with alkoxyorganosilanes carrying different amine substituents.

Materials and methods

The humates from lignite and peat were modified using (3-aminopropyltriethoxy)-silane (APTES) and (1-aminohexamethylenene, 6-aminomethylene)-triethoxysilane (AHATES). The obtained derivatives were characterized using elemental analysis and Fourier transform infrared spectroscopy. Nitrogen release in the form of ammonia or nitrate was evaluated using dissolution tests under sterile aqueous conditions as well as long-term soil experiments. Ammonium and nitrate were determined using ion-selective electrodes. Activity index (AI) was calculated from the dissolution tests. For soil trials, arable Retisol was sampled from 0- to 5-cm layer in Yaroslavl region (Russia). The soil experiments were conducted over 78 days using (NH₄)₂SO₄ as an activator of nitrification and 3-amino-1,2,4-triazole as an inhibitor of autotrophic nitrifying bacteria.

Results and discussion

Modification of lignite and peat humates leads to an increase in nitrogen content up to 2 and 4.3 %, respectively, in case of APTES, and up to 3 and 6 %, respectively, in case of AHATES. All humic derivatives gradually released N upon dissolution in water over 6 days up to 51 % of the total N. The AI values ranged from 4 to 13 %. Amendment of soil with the modified humic materials induced an increase in nitrate content resulting from nitrification of released ammonia by soil microflora. This was confirmed by aminotriasole experiments. The nitrogen release occurred slowly: over the first week of incubation, it did not exceed 36–69 % of the total N content. The higher release rate of ammonium nitrogen was observed for CHS-AHATES versus CHS-APTES derivative, whereas no difference was seen between the two peat derivatives, which showed release rate on the level of CHS-AHATES derivative. Positive effect of all modified humic materials lasted over 78 days.

Conclusions

Modification of lignite and peat humates with two aminoorganosilanes carrying one and two nitrogen atoms in the amine substituent brought about twofold to threefold enrichment of the parent humic materials with nitrogen, which was capable of slow release upon incubation in soils. It was released in the form of ammonia and transformed to nitrates by autotrophic nitrifying soil microflora. There was no clear relationship established between structure of amine substituent of organosilane and slow-release properties of the corresponding humic derivatives. The conclusion was met that principal application of aminoorganosilane derivatives of humic substances (HS) is soil structuring, whereas nitrogen-fertilizing capabilities might be considered as beneficial added-value feature of these humic products.

     

Simultaneous effect of sex and dose on efficacy of clove oil,tricaine methanesulfonate, 2‐phenoxyethanol and propofol as anaesthetics in guppies,Poecilia reticulata (Peters)

We studied the simultaneous effect of sex and dose on anaesthesia efficacy to estimate, if possible, the lowest effective dose (LED) for clove oil, tricaine methanesulphonate (MS‐222), 2‐phenoxyethanol (2‐PE) and propofol in mature guppies. LED is the lowest dose needed to reach A5 stage in a mean time of 3 min, with mean recovery (R5) time of 5 min. We used four doses/anaesthetic: 25, 50, 75 and 100 mg/L for clove oil; 120,140,160 and 180 mg/L for MS‐222; 800, 1,000, 1,200 and 1,400 mg/L for 2‐PE, and 7.50, 8.25, 10.00 and 11.25 mg/L for propofol. Each dose was tested on 10 females and 10 males. Morbidity, mortality and behavioural changes were checked on two control groups (10 males and 10 females/group). Sedation (A3), A5 and R5 times were recorded. Significant interactive effect dose*sex on A5 time was found for each anaesthetic agent (p_dose*sex < .05). Except for MS‐222 (p_dose*sex = .284), significant interactive effect dose * sex on R5 time was found (p_dose*sex < .05). A5 time in females tended to be greater than in males, but, in general, R5 times were longer in males. Body size differences between males and females could explain these differences in MS‐222 on A5 time and for clove oil, 2‐PE and propofol on R5 time. No dose simultaneous meet LED′s conditions relating to both A5 and R5 times; therefore the lowest doses inducing A5 in a mean time of 3 min could be a safe guideline for anaesthetic procedure in both male and females.     

Development and testing of a weather-based model to determine potential yield losses caused by potato late blight and optimize fungicide application

Late blight is one of the most important potato diseases. To minimize yield losses, various protective measures are used including fungicide application. Active use of fungicides results in a contamination of the environment. Therefore, crop protection strategies optimizing the number of treatments are of great interest. Using information about late blight development in an experimental potato field recorded over 30 seasons, a simulator to forecast yield losses caused by the disease was developed based on the number of 5-d periods favorable for reinfection of plants during a vegetation season. The simulator was successfully verified using independent data on the disease development from nine unprotected potato fields in the Netherlands and Germany. The average difference between the calculated and real yield losses did not exceed 5%. Using the simulator and weather data for a period of 2007–2017, yield losses were calculated for several areas of the Bryansk, Tambov, and Orenburg Regions of Russia. The results revealed differences in disease development between these regions and may be used to develop recommendations for a frequency of fungicide applications according to the regional risk of epidemics, leading to a significant reduction in fungicide use.     

Antithrombotic Activity of Brewers’ Spent Grain Peptides and their Effects on Blood Coagulation Pathways

Antithrombotic activity of brewers’ spent grain peptides before and after simulated gastrointestinal digestion and their effects on blood coagulation pathways were evaluated. Two hydrolysates were produced using sequential enzymatic systems: alkaline protease + Flavourzyme (AF) and neutral protease + Flavourzyme (PF). Simulation of gastrointestinal digestion of AF and PF hydrolysates was made using porcine pepsin and pancreatin enzymes, obtaining the corresponding digested samples: AFD and PFD, respectively. Peptides were fractionated by ultrafiltration using a 1 kDa cut-off membrane. Hydrolysates had peptides with medium and low molecular weights (2100 and 500 Da, respectively), and Glu, Asp, Leu, Ala, and Phe were the most abundant amino acids. Gastrointestinal digested hydrolysates presented high proportion of small peptides (~500 Da), and higher amount of Val, Tyr, and Phe than hydrolysates. Mass spectrum (HDMS Q-TOF) of AFD-ultrafiltered fraction <1 kDa exhibited peptides from 500 to 1000 Da, which are not present in AF. PFD showed the generation of new peptides from 430 to 1070 Da. All samples showed thrombin inhibitory activity. However, no effect was observed on prothrombin time. Peptides <1 kDa from hydrolysates and digested samples delayed thrombin and thromboplastin time respect to the control (~63%). Also the samples showed thrombin inhibitory activity at common pathway level. Thus, brewers’ spent grain peptides exerted their antithrombotic activity by inhibiting the intrinsic and common pathways of blood coagulation. This is the first report to demonstrate that brewers’ spent grain peptides are able to delay clotting time after simulated gastrointestinal digestion.     

Assessing the Potential of Rhizobacteria to Survive under Phenanthrene Pollution

Rhizobacteria possess a wide variety of qualities governing their pollutant-catabolic and rhizospheric competences. We investigated how the abilities to degrade phenanthrene and other polycyclic aromatic hydrocarbons (PAHs), to synthesize surfactants and the phytohormone indole-3-acetic acid (IAA), to be motile, and to perform chemotaxis toward phenanthrene and some potential root-exudate components were manifested in rhizobacteria isolated from oil-polluted sites. We observed that most of the examined rhizobacteria had the abilities under consideration and that in some strains, these were strongly affected by the bacterial environment. Only one strain—Sinorhizobium meliloti P221—exhibited increased PAH-degrading, surfactant-producing, and IAA-synthesizing activities, as well as distinct behavioral responses. We conclude that S. meliloti P221 can be used as a model to assess the contributions of all these activities to plant-inoculation-induced reduction in the soil PAH contents. This strain also may be useful for phytoremediation applications.     

Effect of paratuberculosis on the diagnosis of bovine tuberculosis in a cattle herd with a mixed infection using interferon-gamma detection assay

Interferon-gamma (IFN-gamma) detection assay is being applied as an ancillary test to tuberculin tests in the diagnosis of bovine tuberculosis to detect the maximum number of infected animals. Among possible factors influencing the performance of tuberculosis-diagnostic tests, paratuberculosis, a widespread disease in Spain and other European countries, has been pointed out as a cause of false positive reactions. Still, its effect on the sensitivity of these tests in cattle has yet to be fully characterized. The impact of paratuberculosis in the apparent sensitivity of IFN-gamma assay was studied in a bullfighting cattle herd with a mixed tuberculosis-paratuberculosis infection, using culture of Mycobacterium bovis and Mycobacterium avium paratuberculosis as the gold standard to determine the infection status of every animal. A total of 218 animals were slaughtered and sampled for bacteriology after blood sampling. IFN-gamma assay showed a lower apparent sensitivity in animals with a mixed infection (50%) compared to all animals suffering tuberculosis (78.3%). This finding indicates that the presence of paratuberculosis in tuberculosis-infected herds could imply a serious impairment in the sensitivity of IFN-gamma detection test.     

Evaluation of Egg White Protein-Based Coatings to Improve the Protection of Frozen Atlantic Salmon (Salmo salar)

ABSTRACT

The effects of different egg white protein coatings on Atlantic salmon quality preservation after 4 months of frozen storage were evaluated. These coatings increased the yield of the fillets and protected them from lipid oxidation, providing better protection than glazing. Egg white protein coatings were better options than glazing to preserve salmon color during frozen storage. The best results were obtained with the application of coatings before freezing. No clear effects of the type and concentration of plasticizer in most quality parameters were found. Egg white protein + glycerol 2:1 applied before freezing was the best coating for frozen Atlantic salmon protection.     

Development of real-time PCR systems based on SYBR? Green I and TaqMan? technologies for specific quantitative detection of Phoma tracheiphila in infected Citrus

Real-time PCR assays based on SYBR? Green I and TaqMan? technologies were developed for in planta detection and quantification of Phoma tracheiphila, the mitosporic fungus causing ‘mal secco’ disease on citrus. Primers and a hybridization probe were designed on the basis of the internal transcribed spacer (ITS) region of the nuclear rRNA genes. The real-time PCR assays were compared with a classic isolation method in two separate experiments carried out on 6 and 24 month-old sour orange seedlings, artificially inoculated with a conidial suspension of the pathogen. Both technologies made it possible to follow the progression of infection by P. tracheiphila, enabling detection and quantification of the target fungus prior to the development of symptoms. The detection limit was 10 copies of the cloned target sequence and 15 pg of genomic DNA extracted from fungal spores. The values of the cycle threshold (Ct) were linearly correlated with the concentration of the target DNA, indicating that the method is suitable as a qualitative and quantitative assay. The presence of non-target fungal DNA had no effect on the specificity of the assay, but resulted in a 10-fold reduction of sensitivity. Total inhibition of the reaction occurred when conidia of the target pathogen were mixed with an organic soil substrate before extracting DNA using the standard protocol, while an alternative purification kit resulted in a significant decrease in sensitivity. Compared to classic methods, real-time PCR proved faster and easier to perform and showed a higher sensitivity. These results suggest that real-time PCR, based on both chemistries, has a great potential for early diagnosis of ‘mal secco’ disease and for quantitative estimation of fungal growth within host tissue.     

Construction of a recombinant strain of <Emphasis Type="Italic">Pseudomonas fluorescens</Emphasis> producing both phenazine-1-carboxylic acid and cyclic lipopeptide for the biocontrol of take-all disease of wheat

Four Neofabraea species are responsible for bull’s eye rot, which is an important postharvest disease of apples and pears. The species diversity of its causal agents in Europe has not been thoroughly explored using molecular genetic methods. Eighty-one Neofabraea isolates were obtained mostly from apples with bull’s eye rot symptoms in the Czech Republic over a two year period. The isolates were identified using PCR fingerprinting and DNA sequencing of the ITS rDNA region, the mitochondrial SSU rDNA and the β-tubulin and EF1α genes. The most common species was N. alba (89 %), followed by N. perennans (5 %) and N. kienholzii (5 %). This is the third published record of N. kienholzii in Europe. The species identity of the isolate CPPF507, which was placed close to N. kienholzii, remains unclear. EF1α was shown to be a suitable marker for the identification of species of the genus Neofabraea and was comparable to the previously used β-tubulin gene. Furthermore, the aggressiveness of individual species was compared and species distribution across Europe was summarized. N. perennans and isolate CPPF507 proved to be the most aggressive, whereas the least aggressive was N. kienholzii. Two N. alba isolates isolated from symptomless apple fruits and leaves were pathogenic to apples in the infection tests.