11 research outputs found

    Accuracy improvement and standardization on the method of mosquito monitoring and procedure for detection of flaviviruses and bunyaviruses

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    This publication provides information on the methods of mosquito monitoring and detection of viruses. Capturing of mosquitoes – vectores of many diseases of mans and domestic animals depends on the type of trap and attractant used, another important factor is the height at which the trap is situated. Classical methods for arboviruses detection (suckling mouse brain inoculation, CPE in cell culture, etc.) are laborious and are biased towards viruses that grow well in vitro or in vivo. Thus, these classical methods are not always suitable for large-scale surveillance. PCR-based techniques have been shown to be efficient and sensitive in surveillance for individual virus species. We hope that this guide described methods of monitoring of mosquitoes and detection of viruses by RT-PCR will be helpful for the authority of protection public health. Use of these methods enable optimization of mosquitoes eradication by methods economically advantageous and saving to the nature

    Study of biodiversity and account of haematophagous dipterous insects, mainly mosquitoes, in PLA Palava and Litovelské Pomoraví

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    Changes in the biodiversity of haematophagous insects, mainly mosquitoes and effect of artificial flood on the account of mosquitoes were studied in PLA Palava. Global changes of temperature on the biodiversity of mosquitoes in PLA Palava and Litovelské Pomoraví were studied too

    RAP-PCR fingerprinting reveals time-dependent expression of development-related genes following differentiation process of Bacillus thuringiensis

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    Gene expression profiles are important data to reveal gene functions putatively involved in crucial biological processes. RNA arbitrarily primed-polymerase chain reaction (RAP-PCR) and specifically primed reverse transcription PCR (RT-PCR) were combined to screen differentially expressed genes following development of a commercial Bacillus thuringiensis (Bt) subsp. kurstaki strain 8010 (serotype 3a3b). Six differentially expressed transcripts (RAP1 to RAP6) were obtained. The RAP1 encoded a putative triple helix repeat-containing collagen or an exosporium protein H related to spore pathogenicity. The RAP2 was homologous to a ClpX protease and an ATP-dependent protease La (LonB) likely acted as virulence factors. The RAP3 was homologous to a beta subunit of propionyl-CoA carboxylase (PCC) enzyme (PCCB) required for the development of Myxococcus xanthus. The RAP4 had homology to a quinone oxidoreductase involved in electron transport and ATP formation. The RAP5 showed significant homology to a uridine kinase (UDK) mediated phosphorylation of uridine, azauridine. The RAP6 shared high sequence identity with 3-methyl-2-oxobutanoate-hydroxymethyltransferase (MOHMT) (EC 2.1.2.11) [also known as ketopantoate hydroxymethyltransferase (KPHMT) or PanB] involved in the operation of the tricarboxylic acid cycle. The findings described here would help to elucidate the molecular mechanisms underlying the differentiation process of Bt and unravel novel pathogenic genes.The accepted manuscript in pdf format is listed with the files at the bottom of this page. The presentation of the authors' names and (or) special characters in the title of the manuscript may differ slightly between what is listed on this page and what is listed in the pdf file of the accepted manuscript; that in the pdf file of the accepted manuscript is what was submitted by the author

    Mosquito cell line C6/36 shows resistance to Cyt1Aa6

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    265-269This study aimed to investigate the resistance mechanism of C6/36 cells to Cyt1Aa6 protein under selection pressure. Receptor binding properties of Cyt1Aa6 toward sensitive and resistant C6/36 cells were investigated. More sensitive cells were detected with goat-anti-rabbit-FITC-labeled antibody, and the quantity of in vitro activated Cyt1Aa6 toxin bound to resistant cells was greatly reduced. Ligand western blot assays showed that disappearance of the 26 kDa protein and weakness of the positive bands of 68 kDa from resistant cells might lead to the resistance of C6/36 cells to Cyt1Aa6 toxin. The resistance of C6/36 cells was detected under selection in vitro-activated Cyt1Aa6 toxin. Receptor binding demonstrated that reduced Cyt1Aa6 bound to resistant cells, which might be closely related to the disappearance and weakness of some proteins. The results presented here are the first to demonstrate that Cyt1Aa protein, a uniquely characteristic toxin, induced resistance at the cellular level. It might be attributed to the change of receptors

    Cloning, expression and characterisation of a promising mosquitocidal gene

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    799-803A new mosquitocidal gene, cyt1Aa6, was isolated and cloned from the novel Bacillus thuringiensis strain LLP29, previously isolated from the phylloplane of Magnolia denudata. Nucleotide sequence analysis of cyt1Aa6 indicated that the open reading frame consisted of 750 base pairs, encoding 249 amino acid sequences with a calculated molecular weight of 27.3681 kDa and a PI value of 4.77. An homological comparison revealed that the cyt1Aa6 amino acid sequence was 99% identical with those of known Cyt1Aa proteins. In addition, the cyt1Aa6 gene was successfully expressed in Escherichia coli BL21. Bioassays on Aedes albopictus showed that Bt LLP29 and the expressed BL21 were both toxic to 3rd-instar mosquito larvae. The isolation of cyt1Aa6 provides new opportunities for selecting new strains and to obtain novel B. thuringiensis products based on its toxins

    Characterization of cry1Cb3 and cry1Fb7 from Bacillus thuringiensis subsp. galleriae

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    Two cry1-type genes encoding insecticidal crystal proteins (ICPs) were detected by PCR-RFLP and cloned from Bacillus thuringiensis subsp. galleriae 87. The nucleotide sequences were deposited in GenBank with accession numbers EU679501 and EU679502, and designated as cry1Fb7 and cry1Cb3 respectively by B. thuringiensis Delta- Endotoxin Nomenclature Committee. cry1Cb3 shared 99% homology with other cry1Cb genes. The existence of two additional stop codons indicated cry1Cb3 was a silent gene. The cry1Cb3 was 3531 bp with 38.98% G+C content and its first open reading frame (ORF) encoded a protein of 213 amino acid residues with a calculated molecular weight of 23.8 kDa and a predicted pI value of 4.63. Five amino acid sequence blocks (block 1, block 2, block 3, block 4 and block 5) were found in Cry1Cb3. Translation of cry1Fb7 revealed an ORF of 3525 bp with 39.12% G+C content and a protein with a calculated molecular weight of 133.2 kDa and a predicted pI value of 5.18. Cry1Fb7 had five amino acid sequence blocks (blocks 1, 2, 3, 4 and 5) and three domains (I, II and III), which consisted of 218 residues (Leu34 to Ala252), 197 residues (Thr257 to Asp454), and 138 residues (Ile464 to Glu600), respectively
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