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Keywords RNA egzosom
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Abstract

The Bulletin of the Polish Academy of Sciences: Technical Sciences (Bull.Pol. Ac.: Tech.) is published bimonthly by the Division IV Engineering Sciences of the Polish Academy of Sciences, since the beginning of the existence of the PAS in 1952. The journal is peer‐reviewed and is published both in printed and electronic form. It is established for the publication of original high quality papers from multidisciplinary Engineering sciences with the following topics preferred: Artificial and Computational Intelligence, Biomedical Engineering and Biotechnology, Civil Engineering, Control, Informatics and Robotics, Electronics, Telecommunication and Optoelectronics, Mechanical and Aeronautical Engineering, Thermodynamics, Material Science and Nanotechnology, Power Systems and Power Electronics.

Journal Metrics: JCR Impact Factor 2018: 1.361, 5 Year Impact Factor: 1.323, SCImago Journal Rank (SJR) 2017: 0.319, Source Normalized Impact per Paper (SNIP) 2017: 1.005, CiteScore 2017: 1.27, The Polish Ministry of Science and Higher Education 2017: 25 points.

Abbreviations/Acronym: Journal citation: Bull. Pol. Ac.: Tech., ISO: Bull. Pol. Acad. Sci.-Tech. Sci., JCR Abbrev: B POL ACAD SCI-TECH Acronym in the Editorial System: BPASTS.

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Authors and Affiliations

Andrzej Dziembowski
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Abstract

Prof. Andrzej Dziembowski of the PAS Institute of Biochemistry and Biophysics, laureate of this year’s Prize of the Foundation for Polish Science (FNP), talks about RNA-degrading enzymes, the role of yeast in studies that help humans, and two different types of scientists.

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Authors and Affiliations

Andrzej Dziembowski
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Abstract

In this study the quality of total RNA, isolated from fresh spermatozoa, was compared between boars with good and poor semen freezability (GSF and PSF, respectively). Semen from 3 boars with GSF exhibited significantly higher total motility, mitochondrial function, plasma membrane integrity and reduced lipid peroxidation compared with 3 boars with PSF after cryo- preservation. There were variations in the quality of RNA isolated from spermatozoa of boars with GSF and PSF. Boars with GSF exhibited mainly full-length, intact RNA, whereas substantial amounts of degraded RNA were detected in spermatozoa from boars with PSF. Further under- standing of the biological relevance of RNAs in sperm function is critical to improve the freezabil- ity of boar semen.

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Authors and Affiliations

L. Fraser
P. Brym
M. Mogielnicka-Brzozowska
K. Wasilewska
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Abstract

RNA extraction involves several main stages, regardless of the method of extraction: homogenization, effective denaturation of proteins from RNA, inactivation of ribonuclease and removal of any DNA, protein, and some residual contamination. Isolation of undamaged intact RNA is challenging when the related tissue contains high levels of polysaccharides and phenols. Several efforts have been made towards the comparison and optimization of extraction and purification methods for RNA from plant tissues. This is dictated by the necessity of obtaining RNA of a good quality and in a sufficient quantity for further molecular analyzes. Plant storage organs (such as bulbs or seeds) rich in polysaccharide and polyphenolic compounds present distinct challenges for total RNA isolation. Such components, considered in this case as contamination, may bind and co-precipitate with nucleic acids and negatively affect later assays. Since standard routine protocols yield unacceptable results in bulbs, we have designed a new method for RNA extraction. We used two modified procedures (based on CTAB and sarkosyl reagents) of RNA extraction from so called “difficult plant material” and compared them to a popular RNA isolation base on the column isolation kit and TriPure reagent. Our modified protocols dealt with problems of both RNA degradation and low yield caused by co-purification with polysaccharides present in plant bulbs. In this study we have shown that improvement of the CTAB and sarkosyl method with a lyophilization step of plant tissues leads to isolation of high quality RNA from difficult material like storage organs of bulbous plants. The main changes in the procedure compared to the previously described methods concerned the different order of lithium chloride and sodium acetate addition, lithium chloride concentration increase and modification of centrifugation conditions. Gel electrophoresis and spectrophotometer analysis confirmed the high quality and integrity of the obtained RNA. The modified procedures allowed for obtaining a satisfying amount of RNA concentration in the range from 280 to 950 ng/μl depending on the plant species. Thus, the demonstrated RNA isolation methods are efficient and can be used for plant material rich in polysaccharides, such as bulbs.

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Authors and Affiliations

Maria Duszyn
Brygida Świeżawska
Mateusz Kwiatkowski
Krzysztof Jaworski
Adriana Szmidt-Jaworska
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Abstract

This study characterizes mycorrhiza helper bacteria (MHB) from selected unpolluted locations as well as subjected to industrial emissions. To determine the species of bacteria isolated from the roots of ectomycorrhizal pine and birch, a method based on the sequence analysis of a 16S rRNA gene was used. The isolated bacteria were initially characterized by available biochemical methods and phenotypic observation. On the selected bacteria representatives isolation of DNA was performed, on which the PCR reaction was carried out. In this way amplified samples were automatically sequenced and the obtained results were compared to public databases. Among the isolated bacteria Pseudomonas fluorescens SBW25 and Burkholderia xenovorans LB400 species were dominant.

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Authors and Affiliations

Anna Ząbkiewicz
Magdalena Myga-Nowak
Katarzyna Bandurska
Justyna Paczyńska
Anna Szybecka
Piotr Krupa
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Abstract

Nitritation, the first stage of ammonia removal process is known to be limiting for total process performance. Ammonia oxidizing bacteria (AOB) which perform this process are obligatory activated sludge habitants, a mixture consisting of Bacteria, Protozoa and Metazoa used for biological wastewater treatment. Due to this fact they are an interesting bacterial group, from both the technological and ecological point of view. AOB changeability and biodiversity analyses both in wastewater treatment plants and lab-scale reactors are performed on the basis of 16S rRNA gene sequences using PCR-DGGE (Polymerase Chain Reaction – Denaturing Gradient Gel Electrophoresis) as a molecular biology tool. AOB researches are usually led with nested PCR. Because the application of nested PCR is laborious and time consuming, we have attempted to check the possibility of using only first PCR round to obtain DGGE fingerprinting of microbial communities. In this work we are comparing the nested and non-nested PCR-DGGE monitoring of an AOB community and presenting advantages and disadvantages of both methods used. The experiment revealed that PCR technique is a very sensitive tool for the amplification of even a minute amount of DNA sample. But in the case of nested-PCR, the sensitivity is higher and the template amount could be even smaller. The nested PCR-DGGE seems to be a better tool for AOB community monitoring and complexity research in activated sludge, despite shorter fragments of DNA amplification which seems to be a disadvantage in the case of bacteria identification. It is recommended that the sort of analysis approach should be chosen according to the aim of the study: nested-PCR-DGGE for community complexity analysis, while PCR-DGGE for identification of the dominant bacteria.
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Authors and Affiliations

Aleksandra Ziembińska-Buczyńska
Jarosław Wiszniowski
Sławomir Ciesielski
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Abstract

The current study is the first phylogenetic and secondary RNA structure analysis of Dactylogyrus species parasitising gill filaments of Iraqi cyprinid fishes. Most previous phylogenetic studies have targeted on primary DNA sequence data. Nevertheless, RNA secondary configuration is principally helpful in systematics since they comprise features that do not appear in the primary sequence and provide morphological information. The primary objective was molecular-based identification of Dactylogyrids species using evolutionary tree and secondary RNA structure prediction. A total of 681 fish were collected from the Lesser Zab River in the northeast of Iraq in the sub-district of Altun-Kopru from August 2016 to September 2017 and brought to the Zoology Research Laboratory, Salahaddin University-Erbil, Iraq. All fish were classified as 18 cyprinid species. The species of Dactylogyrus were identified by the 28S rDNA subunit using PCR and sequencing methods, and the obtained nucleotide sequences were then compared with the available GenBank sequences. Phylogenetic relationships were concluded using Neighbour-Joining (NJ), Maximum Likelihood (ML), and Minimum Evolution (ME) methods. The results justify the validation of 11 Dactylogyrus species (three of them were newly recorded in Iraq). Additionally, out of nine infected fish species, seven of them were regarded as a new host for Dactylogyrus species. Secondary RNA configuration prediction using minimum free energy was considered as a hopeful tool for species identification. This was considered the first comprehensive phylogenetic study in the area. It was concluded that PCR sequencing, phylogenetic and secondary RNA analysis were proper molecular methods for identifying Dactylogyrids species on the gills of fishes.

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Authors and Affiliations

Q.M.K. Koyee
S.M.A. Abdullah
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Abstract

A filamentous benthic cyanobacteria, strain USMAC16, was isolated from the High Arctic Svalbard archipelago, Norway, and a combination of morphological, ultrastructural and molecular characterisation (16S rRNA gene sequence) used to identify to species level. Cell dimensions, thylakoid arrangement and apical cell shape are consistent with the Pseudanabaena genus description. The molecular characterisation of P. catenata gave 100% similarity with Pseudanabaena catenata SAG 1464-1, originally reported from Germany. Strain USMAC16 was cultured under a range of temperature and photoperiod conditions, in solid and liquid media, and harvested at exponential phase to examine its phenotypic plasticity. Under different culture conditions, we observed considerable variations in cell dimensions. The longest cell (5.91±0.13 μm) was observed at 15°C under 12:12 light:dark, and the widest cell (3.24±0.06 μm) at 4°C under 12:12 light: dark in liquid media. The study provides baseline data documenting the morphological variation of P. catenata in response to changing temperature regimes.
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Authors and Affiliations

Jerzy Smykla
Zoya Khan
Wan Maznah Wan Omar
Faradina Merican Mohd Sidik Merican
Asmimie Asmawarnie Azizan
Choon Pin Foong
Peter Convey
Nazalan Najimudin
Siti Aisyah Alias
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Abstract

An outstanding Polish biochemist, laureate of the Foundation for Polish Science Prize in 2007, member of the Polish Academy of Sciences, a head of the Department of Molecular Biomedicine at the Institute of Bioorganic Chemistry, PAS in Poznan

Professor Wlodzimierz Krzyzosiak’s research path led from the structural chemistry of nucleic acids, through molecular genetics and cancer genetics to molecular medicine. In the last years, Professor's scientific activity focused on understanding the role of RNA in the pathogenesis of human neurological diseases caused by the expansion of repetitive sequences. He also developed new methods of experimental therapy for this group of disorders using antisense oligonucleotides and RNA interference technology. He analyzed the factors influencing the microRNA biogenesis and used this knowledge to improve RNA interference technology tools in therapeutic approaches. Overall, Professor Krzyżosiak coauthored more than 130 publications, which have been cited more than 3500 times so far.

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Authors and Affiliations

Marta Olejniczak
Agnieszka Fiszer
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Abstract

Phenol degradation efficiency or Pscndontonas putida PCM2 l 53 free cel ls was experimentally studied. Bacterial cells were acclimatized to phenol what relied on gradually increasing the phenol concentration in the medium. The highest phenol degradation rate was calculated as approximately 15.2 mgdmŁh'. Investigated strain degraded the phenol at the concentration or 400 111g-d111·-' in 24 h. The result or toxicity analysis showed that acclimatized cells orP putida PCM2 l 53 arc able to survive even al as high concentration or phenol as 3000 rng.dm'. The obtained result suggests that the analyzed strain can be used lor cflcciivc treating of high strength phenolic wastewater. Due to resistance of the strain to high phenol concentration it may be applied in biorcmcdiation of exceedingly contaminated sites, especially where dilution or pollutants cannot be implemented.
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Authors and Affiliations

Grzegorz Przybyłek
Sławomir Ciesielski
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Abstract

Polycyclic aromatic hydrocarbons (PAHs) belong to the group of recalcitrants that on reaching wastewater can irreversibly inhibit some sensitive biological processes in activated sludge such as nitrification. This situation leads to wastewater treatment failure due to the influence of these substances on bacteria responsible for important biochemical processes. Observation of the changes in bacterial diversity using molecular tools, such as denaturing gradient gel electrophoresis (DGGE), could be the first step in finding a way of preventing wastewater treatment failure. The aim of this experiment was to monitor bacterial biodiversity in a membrane bioreactor (MBR) dealing with synthetic wastewater contaminated with high concentration of petroleum organic compounds (POCs) and to study the influence of POCs contamination on bacterial changeability in activated sludge. COD removal in investigated membrane bioreactors was at a level of 93%. The organics removal efficiency was not affected by the maximal tested dose of petroleum contamination ( l OOO μl POCs/l of wastewater) and the MBRs wastewater treatment performance was undisturbed. DGGE analysis revealed that the biodiversity fluctuated slightly in control MBR, while in experimental MBR the biodiversity index decreased drastically after adding the highest experimental concentration of POCs. These results suggest that concentrations of POCs at levels from 50 μl/l to 500 μl/l stimulate biodiversity growth, while the concentration I OOO μI POCs/1 of wastewater seems to inhibit the most sensitive processes in wastewater treatment by influencing the bacterial biocenosis.
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Authors and Affiliations

Aleksandra Ziembińska
Sławomir Ciesielski
Jarosław Wiszniowski
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Abstract

Cells of a multicellular organism are genetically identical but differ in structure and function. This heterogeneity is created by several epigenetic mechanisms during the development of the organism. The epigenetic changes- including DNA methylation, histone post-translational modifications, chromatin remodeling and RNA interference have all been shown to control chromatin structure and regulate a plethora of cellular and organismal processes. There is a strong evidence that epigenetics play a crucial role in the development of diseases such as cancer, schizophrenia or metabolic disorders. The epigenetic regulation underlie memory formation or adaptation to external stimuli. The extent to which environmental effects can provoke epigenetic responses represents an exciting area of future research. Here we review the current knowledge about the epigenetic mechanisms and their relation to the human health and disease.

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Authors and Affiliations

Kamila Pawlicka
Patrick Perrigue
Jan Barciszewski
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Abstract

From the naturally infected cucumber plane spherical virus was isolated that mainly on basis of its serological properties has been identified as Tomato black ring virus (TBRV). Using antiserum against TBRV-ED for the specific crapping of virus followed by PCR test (immunocapture-RT-PCR) allowed co distinguish TBRV from related viruses, especially Beet ringspot virus (BRSV). Presence of as many as rwo satellite RNAs should be found as a unique feature of the cucumber isolace.
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Authors and Affiliations

Henryk Pospieszny
Magdalena Jończyk
Natasza Borodynko
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Abstract

The soil microbiome is exposed to technogenic influence during the operation of metal structures. There are quantitative and qualitative changes in the microbiota of the technogenic ecosystem. During the study of the technogenic soil ecosystem (ferrosphere), samples of which were taken in the field (Chernihiv, Ukraine: 51°29’58”N, 31°16’09”E), the presence of corrosively active microbial cenosis was established: sulfate-reducing, denitrifying, iron-reducing (using acetate as the only electron donor, and Fe (III) as the only electron acceptor) and ammonifying bacteria. The predominant representatives of corrosively active groups of bacteria were isolated. They were identified as Bacillus simplex, Streptomyces gardneri, Streptomyces canus (ammonifying bacteria), Fictibacillus sp. (ammonifying bacteria with iron-reducing ability), Anaerotignum (Clostridium) propionicum (organic acid-producing bacteria), Desulfovibrio oryzae (sulfate-reducing bacteria) based on some microbiological, physiological and biochemical, genetic features. Strains of heterotrophic and hemolitotrophic bacteria (individual representatives and their associations) isolated from the technogenic ecosystem can be used in both industrial and technological spheres. The interaction of isolated bacteria in the process of microbial induced corrosion is a prospect for further research.
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Authors and Affiliations

Nataliia Tkachuk
1
Liubov Zelena
2

  1. Department of Biology, T.H. Shevchenko National University “Chernihiv Colehium”, Hetman Polubotok Str. 53, 14013, Chernihiv, Ukraine
  2. Department of Physiology of Industrial Microorganisms, Danylo Zabolotny Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine, Acad. Zabolotny Str. 154, 03143 Kyiv, Ukraine
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Abstract

Resonance assignment remains one of the hardest stages in RNA tertiary structure determination with the use of Nuclear Magnetic Resonance spectroscopy. We propose an evolutionary algorithm being a tool for an automatization of the procedure. NOE pathway, which determines the assignments, is constructed during an analysis of possible connections between resonances within aromatic and anomeric region of 2D-NOESY spectra resulting from appropriate NMR experiments. Computational tests demonstrate the performance of the evolutionary algorithm as compared with the exact branch-and-cut procedure applied for the experimental and simulated spectral data for RNA molecules.

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Authors and Affiliations

J. Błażewicz
M. Szachniuk
A. Wojtowicz
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Abstract

Ammonia-oxidizing bacteria communities were evaluated in a completely mixed, laboratory scale membrane reactor (MBR) working under anoxic conditions for 5 months. The microorganisms in activated sludge were fed a synthetic medium containing 66-150 mg NH4 +-N/l. The age of the activated sludge in MBR was 50 days and the hydraulic retention time (HRT) was 3.3 days. The estimation of the diversity and complexity of the AOB community together with the identification of the dominant bacteria in the activated sludge under anoxic conditions were performed using denaturing gradient gel electrophoresis (DGGE) and DNA sequencing. Molecular analysis of the microbial community carried out with two microbial molecular markers, 16S rRNA gene and amoA gene, suggested that nitrification was led by a Nitrosomonas-like species. In the biocenosis of the investigated bioreactor, oxygen was the crucial selective parameter. The results obtained in this work showed that amoA gene research is more suitable to study the stability and effectiveness of ammonia oxidation. This information emphasizes the necessity of the usage of molecular markers based on functional genes instead of ribosomal ones in order to present the actual state of the process performed in bioreactors. It was also stated that Nitrosomonas -like bacteria are able to perform nitritation even in anoxic environment, that is probably the reason why these bacteria are the most common AOB in different bioreactors.

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Authors and Affiliations

Aleksandra Ziembińska
Sławomir Ciesielski
Anna Raszka
Korneliusz Miksch
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Abstract

Despite the considerable progress that has recently been made in medicine, the treatment of viral infections is still a problem remaining to be solved. This especially concerns infections caused by newly emerging patogenes such as: human immunodeficiency virus, hepatitis C virus or SARS-coronavirus. There are several lines of evidence that the unusual genetic polymorphism of these viruses is responsible for the observed therapeutic difficulties. In order to determine whether some parameters describing a very complex and variable viral population can be used as prognostic factors during antiviral treatment computational methods were applied. To this end, the structure of the viral population and virus evolution in the organisms of two patients suffering from chronic hepatitis C were analyzed. Here we demonstrated that phylogenetic trees and Hamming distances best reflect the differences between virus populations present in the organisms of patients who responded positively and negatively to the applied therapy. Interestingly, the obtained results suggest that based on the elaborated method of virus population analysis one can predict the final outcome of the treatment even before it has started.

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Authors and Affiliations

P. Kędziora
M. Figlerowicz
P. Formanowicz
M. Alejska
P. Jackowiak
N. Malinowska
A. Frątczak
J. Błażewicz
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Abstract

In this short communication describing experiments carried out on the larvae of two insects, Unaspis euonymi Comstock (feeding on Euonymus japonicus Thunb.) and Dynaspidiotus britannicus Newstead (feeding on Laurus nobilis L.), we evaluate for the first time the efficiency of using DNA insecticides in the control of sap-sucking insects, including armored scale insects. Over a period of 10 days, high insect mortality was detected in both U. euonymi and D. britannicus, accompanied by a significant decrease in the concentration of target RNAs. At the same time, no visible changes were observed when the leaves of the host plants were subjected to treatment with DNA insecticides for one month. The results show the high efficiency of DNA insecticides used against hemipteran insect pests. It is noteworthy that the high efficiency of DNA insecticides and their low cost in comparison with RNA preparations provides a safe and extremely promising potential vehicle for the control of sap-sucking insects.

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Authors and Affiliations

Nikita Gal’chinsky
Refat Useinov
Ekaterina Yatskova
Kateryna Laikova
Ilya Novikov
Mikhail Gorlov
Natalya Trikoz
Alexander Sharmagiy
Yuri Plugatar
Volodymyr Oberemok
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Abstract

5.8S ribosomal RNA plays an important role in protein synthesis and eukaryotic ribosome translocation. Contact DNA insecticides based on antisense fragments of 5.8S ribosomal RNA gene of gypsy moth Lymantria dispar L. showed prospective insecticidal activity on its larvae. The most pronounced insecticidal effect was found for antisense fragments 10 and 11 nucleotides long (oligoRIBO-10 and oligoRIBO-11), whereas 12 nucleotides long fragment (oligoRIBO-12) caused the lowest level of insect mortality. This data corresponds to results obtained earlier using rabbit reticulocyte and wheat germ extracts, where maximum inhibition of protein synthesis was observed when a relevant oligomer 10-11 nucleotides long was used, whilst longer chain lengths resulted in reduced inhibition. Using oligoRIBO-11 fragment we have shown penetration of antisense oligonucleotides to insect cells through insects’ exoskeletons. MALDI technique registered the penetration of the oligoRIBO-11 fragment into insect cells after 30 min and a significant response of insect cells to the applied oligonucleotide after 60 min, which indicates not only that the oligonucleotide enters the insect cells, but also the synthesis of new substances in response to the applied DNA fragment. Contact DNA insecticides developed from the L. dispar 5.8S ribosomal RNA gene provide a novel biotechnology for plant protection using unmodified antisense oligonucleotides.

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Authors and Affiliations

Volodymyr V. Oberemok
Kateryna V. Laikova
Refat Z. Useinov
Nikita V. Gal’chinsky
Ilya A. Novikov
Kseniya A. Yurchenko
Mikhail E. Volkov
Mikhail V. Gorlov
Valentina A. Brailko
Yuri V. Plugatar
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Abstract

Obviously, the moment has come in agriculture and forestry when we must decide to gradually abandon (where possible) non-selectively acting chemical insecticides, taking into consideration the overall decrease in the total biomass of insects, especially pollinators, and the increased number of diseases and human deaths directly or indirectly associated with chemical insecticides. Yet with the world facing the rapid growth of human populations, the annual reduction of cultivated areas, and substantial losses from insect pests, most experts believe that no serious alternative to chemical insecticides exists. However, there is definitely room to create more well-tailored chemical insecticides. And there is hope, in the form of effective DNA insecticides able to provide an adequate level of safety for non-target organisms. In this short communication describing experiments carried out on the larvae of Ceroplastes japonicus Green (feeding on Ilex aquifolium Linnaeus), we show for the first time the enormous potential for the use of DNA insecticides in the control of soft scale insects and how they could replace non-selective organophosphate insecticides.

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Authors and Affiliations

Refat Zhevdetovich Useinov
Nikita Gal’chinsky
Ekaterina Yatskova
Ilya Novikov
Yelizaveta Puzanova
Natalya Trikoz
Alexander Sharmagiy
Yuri Plugatar
Kateryna Laikova
Volodymyr Oberemok
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Abstract

Filtration through biologically active carbon (BAC) filters is an effective method of organic matter removal during drinking water treatment. In this study, the microbial community in the initial period of filters’ operation, as well as its role in the organic matter removal were investigated. Research was carried out in a pilot scale on two BAC filters (Filter 1 and Filter 2) which were distinguished by the type of inflowing water. It was observed that the number of heterotrophic plate count bacteria and total microbial activity were significantly higher in water samples collected from Filter 2, which received an additional load of organic matter and microorganisms. Despite the differences in the values of chemical and microbiological parameters of inflowing water, the composition of the microbiome in both filters was similar. The predominant taxon was a bacterium related to Spongiibacter sp. (Gammaproteobacteria) (>50% of relative abundance). In both filters, the efficiency of organic matter removal was at the same level, and the composition and relative frequency of predicted functional pathways related to metabolism determined using PICRUSt (Phylogenetic Investigation of Communities by Reconstruction of Unobserved States Software) at level 3 of KEGG (Kyoto Encyclopedia of Genes and Genomes) Orthology – were also similar. The study demonstrated that a 40-day period of filter operation after filling with virgin granular activated carbon, was sufficient to initiate biofilm development. It was proved, that during the initial stage of filter operation, microorganisms capable of biodegradation of various organic compounds, including xenobiotics like nitrotoluene, colonized the filters
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Authors and Affiliations

Beata Mądrecka-Witkowska
1
ORCID: ORCID
Małgorzata Komorowska-Kaufman
1
ORCID: ORCID
Alina Pruss
1
ORCID: ORCID
Dorota Holc
1
ORCID: ORCID
Artur Trzebny
2
ORCID: ORCID
Miroslawa Dabert
2
ORCID: ORCID

  1. Poznan University of Technology, Institute of Environmental Engineering and Building Installations, Poznań, Poland
  2. Adam Mickiewicz University in Poznań, Faculty of Biology, Poznań, Poland
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Abstract

Strawberry leaves showing leaf blight symptoms were collected from six different farms in Ismailia and Beheira Governorates in Egypt during the 2020–2021 growing season. Eight bacterial isolates, i.e., Pa1, Pa2, Pa3, Pa4 (Ismailia farms) and Pa5, Pa6, Pa7 and Pa8 (Beheira farms) were isolated. A pathogenicity test of bacterial isolates was carried out using detached strawberry leaf technique. All bacterial isolates produced leaf blight disease symptoms. Isolates Pa2 and Pa6 showed the highest pathogenic characteristics with clear symptoms on detached strawberry leaves. The phenotypic, biochemical and physiological characters of the highest pathogenic isolates were confirmed by PCR analysis using 16S rRNA gene. The two bacterial isolates were identified as Pantoea ananatis with similarity of 97.05% with accession number MH_127816.1 (isolate Pa2, Ismailia), while the isolate ( Pa6, Beheira) with similarity of 97.03% with accession number NR_026045.1. The 16S rDNA sequences were deposited in the GenBank nucleotide databases under accession numbers OM258167 and OM279507, respectively. According to the pathogenicity test, morphological and physiological characteristics as well as molecular data (16S rRNA sequencing analysis), this finding is the first report of P. ananatis as a causal agent of strawberry leaf blight disease in Egypt.
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Authors and Affiliations

Tarek Gomaa Abdel-Gaied
1
ORCID: ORCID
Hassan Abd-El-Khair
1
ORCID: ORCID
Maryan Makram Youssef
2
Shereen Abu El-Maaty
3
Maurice Sabry Mikhail
2

  1. Department of Plant Pathology, National Research Centre, Dokki, Giza, Egypt
  2. Department of Plant Pathology, Faculty of Agriculture, Cairo University, Giza, Egypt
  3. Department of Genetics, Faculty of Agriculture, Cairo University, Giza, Egypt
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Abstract

Here we investigate the microbiomes of the soil samples from the Yamal Peninsula (the surroundings of Salekhard city, Russian Federation) using a high-throughput sequencing approach. The main goal was to investigate the impact of mining on soils within the following regeneration, both during the reclamation practice and natural self-growth. Several quarries were studied, engaged in sand, clay and chromatic ores mining. The taxonomic analysis of the soil microbiomes revealed 50 bacterial and archaeal phyla; among the dominant phyla were: Proteobacteria, Actinobacteria, Acidobacteria, Chroloflexi, Gemmatimonadetes, Verrucomicrobia, Planctomycetes, Bacteroidetes, AD3, and Nitrospirae. Compared to the typical tundra soil, which was chosen as a control, the disturbed soils had increased biodiversity and total counts for soil bacteria, archaea, and fungi, especially in the cryosolic horizon. The different mining strategies caused significantly different transformations of soil microbiomes, which was less pronounced for self-growth compared to reclaimed quarries. This isolation of the reclaimed quarry was mainly associated with the increase of the amount of acidobacteria (fam. Koribacteraceae and Acidobacteriaceae and order Ellin6513), some proteobacterial taxa (fam. Syntrophobacteraceae), and Chloroflexi (fam. Thermogemmatisporaceae). The study also revealed bacteria, which tend to be specific for marine tundra environments: gemmatimonadetes from the order N1423WL and Chloroflexi bacteria from the order Gitt-GS-136.

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Authors and Affiliations

Elizaveta Pershina
Ekaterina Ivanova
Anastasia Kimeklis
Alexey Zverev
Arina Kichko
Tatiana Aksenova
Evgeny Andronov
Evgeny Abakumov
ORCID: ORCID

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