Development of a multiplex RT-PCR assay for simultaneous detection of the major viruses that affect rainbow trout (Oncorhynchus mykiss)     

Ana C. A. S. Pinheiro  Enrico Volpe  Donatella Principi  Santino Prosperi  Sara Ciulli 《Aquaculture International》2016,24(1):115-125

The major viral diseases that affect rainbow trout (Oncorhynchus mykiss) are viral haemorrhagic septicaemia, infectious haematopoietic necrosis, infectious pancreatic necrosis and sleeping disease. In the presented study, we developed a multiplex RT-PCR (mRT-PCR) assay for the simultaneous detection of these four rainbow trout viruses in a single assay. The choice of primers was carried out based on the expected size of the fragments, the temperature and time required for the amplification, and the specificity for the target sequence. Firstly, the method was optimised using reference strains of viral haemorrhagic septicaemia virus (VHSV), infectious haematopoietic necrosis virus (IHNV), infectious pancreatic necrosis virus (IPNV) and sleeping disease virus (SDV) cultivated with permissive cell culture lines; subsequently, the method was used for the identification of these viral infections in rainbow trout samples. Twenty-two samples of rainbow trout, clinically suspected of having viruses, were analysed by the developed method to detect the presence of the four viruses, by directly analysing the animal tissues. The mRT-PCR method was able to efficiently detect the viral RNA in infected cell culture supernatants and in tissue samples, highlighting the presence of single infections as well as co-infections in rainbow trout samples. VHSV/SDV and IHNV/SDV co-infections were demonstrated for the first time in rainbow trout. The mRT-PCR method was revealed to be an accurate and fast method to support traditional diagnostic techniques in the diagnosis of major viral diseases of rainbow trout.  相似文献   

Inhibition of human cAMP-phosphodiesterase as a mechanism of the spasmolytic effect of Matricaria recutita L     

Maschi O  Cero ED  Galli GV  Caruso D  Bosisio E  Dell'Agli M 《Journal of agricultural and food chemistry》2008,56(13):5015-5020

Mechanisms underlying the spasmolytic activity of chamomile still remain unclear. Inhibition of cAMP- and cGMP-phosphodiesterases (PDE) is one of the mechanisms operated by spasmolytic drugs. In this study, the effect of chamomile on PDE was investigated. Human platelet cAMP-PDE and recombinant PDE5A1 were assayed in the presence of infusions prepared from sifted flowers and capitula. LC-ESI-MS/MS analysis showed different compositions in infusions made with sifted flowers and capitula. Chamomile inhibited cAMP-PDE activity (IC50 = 17.9-40.5 microg/mL), while cGMP-PDE5 was less affected (-15% at 50 microg/mL). Among the individual compounds tested, only flavonoids showed an inhibitory effect (IC50 = 1.3-14.9 microM), contributing to around 39% of the infusion inhibition; other compounds responsible for cAMP-PDE inhibition still remain unknown. Although experimental evidence supporting the use of chamomile for gastrointestinal minor spasms dates back to the fifties, cAMP-PDE inhibition as a likely mechanism underlying the spasmolytic activity is reported for the first time.  相似文献   

Diversity of heterotrophic aerobic cultivable microbial communities of soils treated with fumigants and dynamics of metabolic, microbial, and mineralization quotients   总被引：1，自引：0，他引：1  

Stefano Mocali  Donatella Paffetti  Giovanni Emiliani  Anna Benedetti  Renato Fani 《Biology and Fertility of Soils》2008,44(4):557-569

A combination of molecular and classical techniques was used to study the composition, structure, diversity, and dynamics of an aerobic heterotrophic cultivable bacterial community isolated from five different soil samples treated with the fumigant agent 1,3-dichloropropene (1,3-D) and further subjected to nitrogen–phosphorous–potassium (NPK) fertigation (F), amendment (C ₂ and C ₄), and NPK fertigation plus amendment (F + C) in two different periods (May and July). The restriction and sequence analysis of 16S rDNA from 189 isolates revealed a very high percentage (94%) of Gram-positive bacterial isolates, most of which (83%) belonging to the genus Bacillus. The degree of intraspecific genetic diversity was high, as shown by random amplified polymorphic DNA analysis. These data seem to be related with the increase in microbial biomass C (C _mic) content and the decrease in the total organic C (C _org) and metabolic quotient (qCO₂) values, especially in amended soils (C ₂, C ₄) where soil microflora mineralized the organic matter of the added fertilizers. In a short term, it is suggested that the presence of very high percentage of Gram-positive bacteria might be related to the ability of these bacteria to form spores so as to be resistant to fumigants rather than being the result of a selective pressure in the predominance of microbial species with a set of genes involved in biodegradation of 1,3-D. Stefano Mocali and Donatella Paffetti contributed equally to this work.  相似文献   

Efficacy of Posidonia oceanica Extract against Inflammatory Pain: In Vivo Studies in Mice     

Laura Micheli  Marzia Vasarri  Emanuela Barletta  Elena Lucarini  Carla Ghelardini  Donatella Degl&#x;Innocenti  Lorenzo Di Cesare Mannelli 《Marine drugs》2021,19(2)

Posidonia oceanica (L.) Delile is traditionally used for its beneficial properties. Recently, promising antioxidant and anti-inflammatory biological properties emerged through studying the in vitro activity of the ethanolic leaves extract (POE). The present study aims to investigate the anti-inflammatory and analgesic role of POE in mice. Inflammatory pain was modeled in CD-1 mice by the intraplantar injection of carrageenan, interleukin IL-1β and formalin. Pain threshold was measured by von Frey and paw pressure tests. Nociceptive pain was studied by the hot-plate test. POE (10–100 mg kg⁻¹) was administered per os. The paw soft tissue of carrageenan-treated animals was analyzed to measure anti-inflammatory and antioxidant effects. POE exerted a dose-dependent, acute anti-inflammatory effect able to counteract carrageenan-induced pain and paw oedema. Similar anti-hyperalgesic and anti-allodynic results were obtained when inflammation was induced by IL-1β. In the formalin test, the pre-treatment with POE significantly reduced the nocifensive behavior. Moreover, POE was able to evoke an analgesic effect in naïve animals. Ex vivo, POE reduced the myeloperoxidase activity as well as TNF-α and IL-1β levels; further antioxidant properties were highlighted as a reduction in NO concentration. POE is the candidate for a new valid strategy against inflammation and pain.  相似文献   

Can a golf course support biodiversity and ecosystem services? The landscape context matter     

Petrosillo  Irene  Valente  Donatella  Pasimeni  Maria Rita  Aretano  Roberta  Semeraro  Teodoro  Zurlini  Giovanni 《Landscape Ecology》2019,34(10):2213-2228

Landscape Ecology - In the last 30 years, the number of golf courses has increased dramatically worldwide. Since no other sport occupies and manages such large areas of green space, landscape...  相似文献   

Antioxidant and Anti-Inflammatory Agents from the Sea: A Molecular Treasure for New Potential Drugs     

Marzia Vasarri  Donatella Degl&#x;Innocenti 《Marine drugs》2022,20(2)

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Collecting horseradish (Armoracia rusticana, Brassicaceae): local uses and morphological characterization in Basilicata (Southern Italy)     

Giulio Sarli  Antonino De Lisi  Rosa Agneta  Sabrina Grieco  Giovanna Ierardi  Francesco Montemurro  Donatella Negro  Vincenzo Montesano 《Genetic Resources and Crop Evolution》2012,59(5):889-899

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Glucoraphasatin and glucoraphenin, a redox pair of glucosinolates of brassicaceae, differently affect metabolizing enzymes in rats     

Barillari J  Iori R  Broccoli M  Pozzetti L  Canistro D  Sapone A  Bonamassa B  Biagi GL  Paolini M 《Journal of agricultural and food chemistry》2007,55(14):5505-5511

Brassica vegetables are an important dietary source of glucosinolates (GLs), whose breakdown products exhibit anticancer activity. The protective properties of Brassicaceae are believed to be due to the inhibition of Phase-I or induction of Phase-II xenobiotic metabolizing enzymes (XMEs), thus enhancing carcinogen clearance. To study whether GLs affect XMEs and the role of their chemical structure, we focused on two alkylthio GLs differing in the oxidation degree of the side chain sulfur. Male Sprague-Dawley rats were supplemented (per oral somministration by gavage) with either glucoraphasatin (4-methylthio-3-butenyl GL; GRH) or glucoraphenin (4-methylsulfinyl-3-butenyl GL; GRE), at 24 or 120 mg/kg body weight in a single or repeated fashion (daily for four consecutive days), and hepatic microsomes were prepared for XME analyses. Both GLs were able to induce XMEs, showing different induction profiles. While the inductive effect was stronger after multiple administration of the higher GRH dosage, the single lower GRE dose was the most effective in boosting cytochrome P-450 (CYP)-associated monooxygenases and the postoxidative metabolism. CYP3A1/2 were the most affected isoforms by GRH treatment, whereas GRE induced mainly CYP1A2 supported oxidase. Glutathione S-transferase increased up to approximately 3.2-fold after a single (lower) GRE dose and UDP-glucuronosyl transferase up to approximately 2-fold after four consecutive (higher) GRH doses. In conclusion, the induction profile of these GLs we found is not in line with the chemopreventive hypothesis. Furthermore, the oxidation degree of the side chain sulfur of GLs seems to exert a crucial role on XME modulation.  相似文献   

Synthetic pyrazole derivatives as growth inhibitors of some phytopathogenic fungi   总被引：3，自引：0，他引：3  

Vicentini CB  Romagnoli C  Andreotti E  Mares D 《Journal of agricultural and food chemistry》2007,55(25):10331-10338

The present study was carried out to investigate the antifungal activity of pyrazole/isoxazole-3-carboxamido-4-carboxylic acids, 4-oxo-5-substituted pyrazolo[3,4-d]pyrimidine-6-thiones, and N-alkyl/aryl-N'-(4-carbethoxy-3-pyrazolyl)thioureas against Pythium ultimum, Botrytis cinerea, and Magnaporthe grisea. The results on growth inhibition showed differences in the sensitivity of the three fungi to the tested substances, and in general P. ultimum was shown to be the most sensitive. On all phytopathogens the best results within the pyrazole/isoxazolecarboxamide series are given by the compounds with the carboxamide and carboxylic groups in positions 3 and 4; the presence of these groups seems to be critical for biological activity in this series of compounds. Among the pyrazolopyrimidines the derivative supplied with the benzylic group was the most active on the three fungi and in particular against P. ultimum. Several compounds belonging to the thiourea series are able to inhibit selectively M. grisea at 50 and 10 microg mL(-1), doses at which the reference commercial compound tricyclazole had low or no effect.  相似文献   

Synthesis of pyrazole derivatives and their evaluation as photosynthetic electron transport inhibitors     

Vicentini CB  Mares D  Tartari A  Manfrini M  Forlani G 《Journal of agricultural and food chemistry》2004,52(7):1898-1906

Two series of new pyrazoles, namely six pyrazolo[1,5-a][1,3,5]triazine-2,4-dione and four pyrazolo[1,5-c][1,3,5]thiadiazine-2-one derivatives, were synthesized as potential inhibitors of the photosynthetic electron transport chain at the photosystem II level. The compounds were confirmed by 1H NMR, elemental, and IR analyses. Their biological activity was evaluated in vivo upon both the growth of blue-green algae and the photosynthetic oxygen evolution by eukaryotic algae and in vitro as the ability to interfere with light-driven reduction of ferricyanide by isolated spinach chloroplasts. Some compounds exhibited remarkable inhibitory properties, comparable to those of the reference commercial herbicides lenacil, diuron, and hexazinone. Results suggest that the substitution of triazine with thiadiazine ring may act as amplifier for herbicidal activity.  相似文献