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EN
Pure tantalum and niobium oxides were produced from the molten cake after mixing oreNH4HF2-KOH in a respective mass ratio of 1/4/3. This resulted in solubilization yields of over 95% for Nb and around 92% for Ta in sulfuric acid solution 3 molar. Extraction of tantalum and niobium using octan-2-ol yielded over 95% niobium and 98% tantalum when pH is set at 0.5-1.0 and 1.5-2 respectively. The compounds K3NbOF6, K3NbO4 and K2TaF7 were identified after melting. While in aqueous solution ionic species such as NbF6 (OH) 23-, NbF6- and TaF72- are likely to be identified. Precipitation of tantalum and niobium in NH4OH solution (pH=7.50-8.0) as hydrated oxides after stripping with distilled water and crystallization identified hydrated oxides such as Ta2O5.nH2O, Nb2O5.nH2O. These two oxides were associated with a small amount of SiO2.nH2O as an impurity resulting from the extraction and precipitation of tantalum and niobium. SEM-EDS, XRD, TGA-DTG and FTIR analyses identified and characterized these compounds.
EN
The genus Scopulariopsis contains over 30 species of mitosporic moulds, which although usually saprophytic may also act as opportunistic pathogens in humans. They have mainly been associated with onychomycosis, and only sporadically reported as a cause of deep tissue infections or systemic disease. Identification of Scopulariopsis species still largely relies on phenotype-based methods. There is a need for a molecular diagnostic approach, that would allow to reliably discriminate between different Scopulariopsis species. The aim of this study was to apply sequence analysis of partial 28S rRNA gene for species identification of Scopulariopsis clinical isolates. Although the method employed did reveal some genetic polymorphism among Scopulariopsis isolates tested, it was not enough for species delineation. For this to be achieved, other genetic loci, within and beyond the rDNA operon, need to be investigated.
EN
In some legal proceedings, the species identification of animal on the basis of fragments of biological material is extremely difficult. This applies both to closely-related and to distant species characterized by similar morphological features. In such circumstances, methods of molecular biology are used, whose evidential value is definitely not in doubt. Histopathological scraps may also have to be used for identifying tests. The aim of the present study was to verify the possibility of using DNA analysis in determining the species of animals on the basis of biological material contained in archival histopathological samples. The examined material consisted of twenty-eight histopathological preparations stained with hematoxylin and eosin. The samples had been prepared from the liver, kidney, spleen, and skeletal muscles. Their age varied from one to seventeen years. Specimens (from twelve species) were identified by inputting sequences in the Barcode of Life Database species identification tool on the basis of the similarity percentage figure from the BOLD report. It was found that genetic tests can effectively identify animal species through the analysis of biological material from histopathological samples.
EN
Molecular phylogeny of crocidurine shrews (Insectivora, Soricidae) in northeastern Asia was investigated to confirm the taxonomic status of unidentified specimens of Crocidura from Cheju Island, South Korea. Phylogenetic trees were constructed by neighbor-joining (NJ) and maximum likelihood (ML) methods, based on mitochondrial cytochrome b gene sequences (402 base pairs) of 37 individuals of seven crocidurine species and three unidentified specimens from 31 localities mainly in northeastern Asia. Phylogenetic position of the three unidentified specimens from Cheju Island were compared with those of Suncus murinus, C. attenuata, C. dsinezumi, C. lasiura, C. sibirica, C. suaveolens, and C. watasei. Both in NJ and ML trees, the three unidentified specimens were included in the cluster of C. dsinezumi and were obviously different from C. suaveolens on Cheju Island. Thus, the present investigation demonstrated that both C. suaveolens and C. dsinezumi exist on Cheju Island.
PL
W artykule omówiono bieżące kierunki badań z dziedziny biologii kryminalistycznej tzw. wildlife forensics, kryminalistyki dzikiej przyrody, w ujęciu badań identyfikacyjnych zwierzęcego DNA gatunków zagrożonych wyginięciem. Przedstawiono zarys problematyki związanej z handlem zwierzętami należącymi do gatunków zagrożonych wyginięciem oraz przytoczono przepisy prawne obowiązujące w tym zakresie. Wymieniono rodzaje śladów i dowodów rzeczowych pochodzenia zwierzęcego, a także sposoby ich zabezpieczania. Pośród różnych rodzajów identyfikacji zwierząt CITES wyróżniono i przedstawiono główne metody genetycznej identyfikacji materiału pochodzenia zwierzęcego gatunków zagrożonych wyginięciem. Osiągnięcia w dziedzinie badań genetycznych z ostatnich dziesięciu lat wskazują na najczęstsze i najbardziej skuteczne zastosowanie w identyfikacji zwierzęcego DNA metod wykorzystujących polimorfizm mitochondrialnego DNA (barkoding DNA, analiza SNP). W przypadkach gdy ocena na podstawie morfologii okazu danego gatunku nie jest możliwa lub dałaby niewiarygodne wyniki badań, zastosowanie molekularnych metod identyfikacji gatunkowej może być bardzo pomocnym narzędziem. Na przykładzie badań przeprowadzonych na wysoko przetworzonych tkankach zwierzęcych wchodzących w skład mieszanek medykamentów tradycyjnej medycyny azjatyckiej przedstawiono rozwiązania umożliwiające identyfikację poszczególnych gatunków zwierząt objętych ochroną. Podsumowaniem artykułu są zalecenia ISFG dotyczące standaryzacji przeprowadzania kryminalistycznych badań DNA pochodzenia zwierzęcego.
EN
The article discusses current research in the field of forensic biology – wildlife forensics in terms of identification of endangered species by DNA analysis. The author provides an overview of illegal trafficking in endangered animal species and relevant legislation. Types of traces and evidence of animal origin and methods of their preservation have been presented. The genetic ways of endangered species identification were shown amongst various methods of CITES identification. The recent decade of DNA research has demonstrated that the use of mitochondrial DNA (mtDNA barcoding, single nucleotide polymorphisms – SNPs) is the best and most effective method of animal DNA identification. In cases where morphological identification of species is not possible or fails to produce reliable results, the application of molecular methods of species identification can be quite helpful. The studies conducted on highly processed animal tissues which are used in traditional Asian medicine products provided solutions for the identification of protected species contained in mixtures. In a final part of the paper, the author presents the ISFG recommendations for standardization in the field of non–human (animal) forensic DNA investigations.
EN
DNA-based identification of species for phylogenetic analysis as well as forensic identification is widely being carried out with the help of polymerase chain reaction (PCR). In this study, a successful effort has been made to identify 5 species of Indian freshwater turtles, including 3 hard-shell turtles (Geoemydidae), i.e. Kachuga dhongoka, K. kachuga and Geoclemys hamiltoni, and 2 species of soft-shell turtles (Trionychidae), i.e. Aspideretes gangeticus and Lissemys punctata punctata, by using a well-optimized PCR-RFLP method. The analysis of nucleotide sequence variations in the PCR-amplified mitochondrial cyt-b genes (encoding cytochrome b) from the 5 species revealed its usefulness in the taxonomic differentiation of these species. On the basis of cyt-b sequence data and the PCR-RFLP pattern, a phylogeny was developed to resolve the genetic relationships between these species, living in the same habitat type. In comparison, the PCR-RFLP of mitochondrial 16S rDNA genes appeared less decisive in analysing phylogenetic relationships or even in species differentiation. Further, the molecular method (PCR-RFLP) developed here is simple, rapid, reliable and reproducible; hence it can be routinely applied for species identification, essential for conservation and management of endangered chelonian species.
EN
DNA barcoding is a practical tool for species identification, when morphological classification of an organism is difficult. Herein we describe the utilisation of this technique in a case of ophthalmomyiasis interna. A 12-year-old boy was infested during a summer holiday in northern Norway, while visiting an area populated with reindeer. Following medical examination, a Diptera larva was surgically removed from the boy’s eye and tentatively identified from its morphological traits as Hypoderma tarandi (L.) (Diptera: Oestridae). Ultimately, DNA barcoding confirmed this impression. The larval cytochrome c oxidase subunit 1 (COI) DNA sequence was matched with both profiles of five adult H. tarandi from the same region where the boy was infested, and other established profiles of H. tarandi in the Barcode of Life Data Systems (BOLD) identification engine.
EN
The aim of this study was molecular identification of S. aureus strains isolated from mastitic milk samples and establishing the genetic relationship between strains isolated from cows belonging to the same herd. In all 43 isolated strains the gap gene (930 bp) was amplified, which enabled their affiliation to the Staphylococcus genus to be established. PCR-RFLP with AluI endonuclease of the gap gene as well as nuc (450 bp) and coa (1130 bp) gene amplification allowed precise S. aureus species identification. One hundred percent of the genetic relationship between strains was established via RAPD-PCR and coa-typing.
EN
It has recently been suggested that goitered gazelles (Gazella subgutturosa and Gazella marica) have paraphyletic maternal origin, and that the mitochondrial cytochrome b gene fragment can be used for species identification prior to reintroduction of the gazelles. Although there is a large geographic area where the gazelles have intermediate morphology, previous researchers have not inferred any signs of mitochondrial haplotype introgression, and it is thought that the introgression, if it exists, is male-biased. We studied mitochondrial haplotypes of morphologically typical G. subgutturosa from two geographic locations. Goitered gazelles from eastern Turkey, morphologically identical to G. subgutturosa, had haplotypes identical to G. marica. This finding confirms ongoing maternal gene introgression from G. marica to G. subgutturosa. Our suggestion is that there is a natural gene flow between these two nominal species, and morphological characters together with recombinant genetic markers rather than mitochondrial DNA should be used to differentiate among individuals from areas close to the contact zone.
EN
The aim of the study was to apply a polymerase chain reaction (PCR) to confirm a presence of hake (genus Merluccius) in a pool of processed seafood samples. A short fragment of the cytb in the mitochondrial control region was amplified. The hake- specific PCR product, due to its limited size, was obtained in a variety of tissue samples. A different level of DNA concentration and degradation, including processed food products, was observed. A total of 64 products were tested and despite their labels, 13 of them were proved to contain no hake. Our protocol for hake identification is faster and more convenient than currently employed methods and may facilitate the identification of fish species.
EN
Electrophoretic methods can be used to identify meat of various animal species. The protein electrophoresis, especially the IEF of the sarcoplasmic proteins, is a wellestablished technique for species identification of raw fish and is used in the control of seafood authenticity. However, in the case of the analysis of heat-processed fish, the method is applicable only to those species which possess characteristic patterns of the heat-stable parvalbumins. Heat-denatured fish muscle proteins may be solubilised by urea or sodium dodecylsulfate (SDS) and separated by urea-IEF or SDS-PAGE, respectively. The comparison of these two methods allowed to conclude that, basically, each of them can be used for species identification of heated fishery products. However, extensively washed products may be preferentially analysed by the SDS-PAGE, because most of the parvalbumins are washed out leaving mainly myosins. On the other hand, the IEF method may be preferred for the differentiation of closely related species rich in parvalbumins isoforms. It is evident from the literature data that species-specific protein separations yield proteins of low molecular weight made up of three light chains of myosin (14-23 kDa), troponin (19-30 kDa) and parvalbumin (about 12 kDa). Investigations showed that the SDS-PAGE method can be used to identify meats of: cattle, sheep, lambs, goats, red deer and rabbits. The technique allowed researchers to identify the following myofibrillar and sarcoplasmic muscle proteins: myosin and actin, -actinin, tropomyosin, troponin. SDS-PAGE allowed the identification of myofibrillar proteins taking into account their molecular weights which was not possible with the assistance of the PAGIF because too many protein bands were obtained. It was possible to obtain differences in the separation of proteins characteristic for certain species, e.g. beef, resulting from the presence of single myofibrillar proteins
PL
Metodami elektroforetycznymi można identyfikować mięso różnych gatunków ssaków i ptaków. Elektroforeza białek, a zwłaszcza IEF białek sarkoplazmy jest dobrze poznaną techniką do identyfikacji gatunków surowych ryb i jest stosowana do kontroli autentyczności produktów morza. Natomiast w wypadku produktów rybnych poddanych procesom obróbki cieplnej metoda ma zastosowanie tylko do oznaczania gatunków dających charakterystyczne rozdziały stabilnych po ogrzaniu parwalbumin. Zdenaturowane w wysokiej temperaturze białka mięśni ryb mogą być rozpuszczone przez mocznik lub SDS i rozdzielone przez zastosowanie odpowiednio IEF lub SDS-PAGE. Porównując obie metody stwierdzono, że w zasadzie są one odpowiednie do identyfikacji gatunkowej ogrzewanych produktów rybnych. Jednak surowce intensywnie myte lepiej analizować metodą SDS-PAGE, ponieważ większość parwalbumin jest wymywana, a zostaje głównie miozyna. Jednocześnie IEF z mocznikiem jest lepszą metodą do różnicowania blisko spokrewnionych gatunków bogatych w izoformy parwalbumin. Z danych literaturowych wynika, że specyficzne gatunkowo rozdziały białek dają białka o małej masie cząsteczkowej złożone z trzech lekkich łańcuchów miozyny (14-23 kDa), troponiny (19-30 kDa) i parwalbuminy (około 12 kDa). Badania wykazały, że metodą SDS-PAGE można różnicować mięso bydła, owiec, jagniąt, kóz, jeleni i królików. Techniką tą identyfikowano szczególnie miofibrylarne białka mięśni: miozynę i aktynę, a-aktininę, tropomiozynę i troponinę. SDS-PAGE pozwoliła na rozpoznanie tych białek z uwzględnieniem ich mas cząsteczkowych, co nie było możliwe z zastosowaniem PAGIF, ponieważ uzyskano zbyt dużą liczbę pasm białek. Stało się możliwe uzyskać różnice w rozdziałach białek charakterystyczne dla pewnych gatunków, np. wołowiny, wynikające z obecności pojedynczych wyróżniających się białek.
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