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1.
A new real-time PCR detection system was developed for grapevine yellows (GY) using TaqMan minor groove binder probes and including two amplicons for group-specific detection of Flavescence dorée (FD) and Bois noir (BN) phytoplasmas, plus a universal phytoplasma amplicon. FD and BN amplicons were designed to amplify species-specific genomic DNA fragments and the universal amplicon to amplify the 16S ribosomal DNA region. Efficiency of PCR amplification, limit of detection, range of linearity and dynamic range were assessed for all three amplicons. The specificity of detection systems was tested on several other isolates of phytoplasmas and bacteria and on healthy field grapevine and insect samples. No cross-reactivity with other phytoplasma strains, plant or insect DNA was detected. The assay was compared with conventional PCR on more than 150 field grapevine, insect and field bindweed samples. Real-time PCR showed higher sensitivity as phytoplasmas were detected in several PCR-negative and in all PCR-positive samples. A data-mining analysis of results from both detection approaches also favoured real-time PCR over conventional PCR diagnostics. The developed procedure for detection of phytoplasmas in grapevine also included amplification of plant DNA co-extracted with phytoplasmic DNA, providing additional quality control for the DNA extraction and PCR amplification for each sample. The newly developed assay is a reliable, specific and sensitive method easily applicable to high-throughput diagnosis of GY. 相似文献
2.
Mycoplasma gallisepticum is a major pathogen of poultry. Mycoplasma imitans is genetically and antigenically closely related to M. gallisepticum, but so far, only a few proteins of M. imitans have been identified as sharing epitopes with M. gallisepticum. In this study, we identified three proteins of M. gallisepticum that share with M. imitans epitopes defined by monoclonal antibodies (MAbs). MAb 9D4 reacted with the 67-kD hemagglutinin V1hA (previously termed pMGA) of M. gallisepticum and with its continuously expressed 40-kD protein. This MAb also reacted with a 40-kD protein of M. imitans, but not with its putative V1hA. Two-dimensional (2D) immunoblots of M. gallisepticum strains showed that their 40-kD proteins reacting with MAb 9D4 are expressed as major forms with isoelectric points (pI) around 6, and also as less-abundant forms differing in pI. In M. imitans, major forms of 40-kD proteins recognized by MAb 9D4 had pI around 6, whereas minor forms had pI between 5.5 and 5.8. The N-terminal sequence of the M. gallisepticum 40-kD protein recognized by MAb 9D4 strongly indicates that this protein is pyruvate dehydrogenase E1, subunit alpha (PdhA protein, also termed AcoA). The position of elongation factor Tu (EF-Tu), detected by the reference MAb GB8, was very similar in the 2D proteome maps of M. gallisepticum and M. imitans (MW of about 45 kD; pI - 5.6). In both M. gallisepticum and M. imitans, MAb 7G1 reacted with proteins of about 36 kD with similar charges (major forms with pI of about 8). The position of this protein in the proteome map of M. gallisepticum and its N-terminal sequence strongly suggest that MAb 7G1 recognizes lactate (malate) dehydrogenase (Ldh or Mdh). Comparison of 2D proteomes of 10 M. gallisepticum strains indicated that positions of EF-Tu, PdhA, and Ldh proteins are rather consistent and can be used as reference points in further analyses of the M. gallisepticum proteome. 相似文献
3.
Steen Lykke Nielsen Annie Enkegaard Mogens Nicolaisen Per Kryger Mojca Vir??ek Marn Irena Mavri? Ple?ko Andreas Kahrer Richard A. Gottsberger 《European journal of plant pathology / European Foundation for Plant Pathology》2012,133(3):505-509
Experiments were carried out to investigate whether Potato spindle tuber viroid (PSTVd) can be transmitted intra- and inter-species from infected Solanum jasminoides to non-infected S. jasminoides and S. esculentum and from infected Brugmansia sp. to S. esculentum by Frankliniella occidentalis and Thrips tabaci by leaf sucking. The F. occidentalis experiments also included feeding on pollen prior to feeding on PSTVd-infected leaf. No thrips-mediated transmission of PSTVd was recorded. The possibility of PSTVd transmission by Apis mellifera and Bombus terrestris during their feeding/pollinating activities within ornamentals and from ornamentals to S. esculentum was included, and no bee-mediated transmission was recorded. 相似文献
4.
Food is physically manipulated by other materials during production processes, and therefore, food quality and safety are vital in processes where foods are in contact with various materials. Wooden frames were used for centuries for dried egg pasta trays; however, with the development of different materials, wood was slowly abandoned and replaced by plastic. Nevertheless, there are some hygienic considerations concerning plastic frames in the dried egg pasta making industry. In this study, plastic and wooden trays were analysed by swabbing (n?=?210) and evaluated in regard to total number of aerobic counts (TAC), Enterobacteriaceae, Escherichia coli, moulds, yeasts and Staphylococcus aureus using dry medium plates. The aims of the research were to (1) evaluate the total number of microorganisms on wood and plastic material used for pasta trays and (2) make a hygienic evaluation of analysed materials for application in the pasta industry. The research was aimed to answer the question, ‘Does the tray material and/or location of the swab sample influence the colony forming unit (CFU)/20?cm2?’ Results showed a statistical difference in CFU/20?cm2 for all bacterial determinations, except for E. coli which was not detected in swabs taken from wooden or plastic trays. This hygiene evaluation study supported the conclusion that the use of wood is appropriate in the food industry from a hygienic and technological point of view. 相似文献
5.
Lunder Mojca Roškar Irena Hošek Jan Štrukelj Borut 《Plant foods for human nutrition (Dordrecht, Netherlands)》2019,74(1):47-53
Plant Foods for Human Nutrition - The diet rich in fruits and vegetables reduces the risk of metabolic syndrome, including diabetes development by various mechanisms of action, mainly due to the... 相似文献
6.
Stanislav Trdan Jana Čuk Anka Poženel Mojca Bavcon Kralj Mojca Rot Branko Carlevaris 《Acta Agriculturae Scandinavica, Section B - Plant Soil Science》2019,69(2):174-180
Between 2012 and 2014, we tested the efficacy of different synthetic attractants for the purpose of massive trapping of common European cockchafer adults (Melolontha melolontha). The research took place in three different locations in Slovenia (Otlica, ?rni Vrh nad Idrijo, Cesta nad Ajdov??ino) during flight periods of adult beetles. In the period 2013–2014, we used, on the basis of the preliminary test results (2012), the following chemicals: toluquinone, cis–3–hexen–1-ol, ethyl acetate, toluquinone?+?cis–3 hexen–1-ol, and ethanol as a control. M. melolontha adults were most abundant in the location ?rni Vrh nad Idrijo; the traps in this location caught 18 times more of them than those in the first location and more than six times more than those in the third location. Male insects accounted for 73 to 87% of the trapped specimens. We confirmed the highest efficiency of cis–3–hexen–1-ol in 2013 (43.25?±?0.08 males/trap), as well as in 2014 (15.00?±?0.14 males/trap). On the basis of the trapped adult common European cockchafers and the simple economic analysis of applying different synthetic substances for attracting them, we found that independent application of cis-3-hexen-1-ol is the most efficient and cost-effective option for attracting the studied insect pest. Consequently, we recommend it as an attractant in the traps for massive trapping of adult common European cockchafers. 相似文献
7.
Bercic RL Slavec B Lavric M Narat M Bidovec A Dovc P Bencina D 《Veterinary microbiology》2008,127(1-2):147-154
Mycoplasma synoviae isolates differ in patterns of immunogenic proteins, but most of them have not been identified yet. The main aim of this study was their identification in two closely related M. synoviae isolates, ULB 02/P4 and ULB 02/OV6, recovered recently from chickens in Slovenia. N-terminal sequencing identified 17 M. synoviae proteins. Amongst them were 14 major, highly expressed but previously unidentified proteins, including enzymes, chaperones and putative lipoproteins. ULB 02/P4 proteins with increasing molecular weight (M(w)) in the region above the lipoprotein MSPB (approximately 40 kDa) were elongation factor EF-Tu, enolase, NADH oxidase, haemagglutinin MSPA, ATP synthase beta chain, trigger factor, pyruvate kinase and chaperone DnaK. Enolase (approximately 47 kDa) seemed to be immunogenic for chickens infected with M. synoviae, whereas EF-Tu, which might cross-react with antibodies to the P1 adhesin of Mycoplasma pneumoniae, was not. ULB 02/OV6 synthesized several immunogenic proteins and those with M(w) of approximately 70, 78, 82, 90, 110 and 160 kDa, cross-reacted with antibodies to Mycoplasma gallisepticum. They remain to be identified, because besides putative lipoproteins, protein bands of 78, 82, 85 and 110 kDa contained also dehydrogenase PdhD, elongation factor EF-G, enzyme PtsG and putative neuraminidase, respectively. 相似文献
8.
Thibaut Olivier Vaidevutis Šveikauskas Elisabeth Demonty Kris De Jonghe Pascal Gentit Mojca Viršček-Marn Sabine Grausgruber-Gröger Sébastien Morio Francesco Faggioli Michèle Visage Frédéric Fauche Maria Gusina Marta Luigi Helena Lasner Irena Mavrič-Pleško 《European journal of plant pathology / European Foundation for Plant Pathology》2016,144(3):645-654
9.
Kropf U Golob T Nečemer M Kump P Korošec M Bertoncelj J Ogrinc N 《Journal of agricultural and food chemistry》2010,58(24):12794-12803
Isotope parameters (δ(13)C(honey), δ(13)C(protein), δ(15)N) were determined for 271 honey samples of 7 types (black locust, multifloral, lime, chestnut, forest, spruce, and fir honeys) from 4 natural geographical regions of Slovenia. Carbon and nitrogen stable isotope ratios were measured to elucidate the applicability of this method in the identification of the botanical and geographical origin of honey and in honey adulteration. Only 2.2% of the samples were adulterated according to the internal standard carbon isotope ratio analysis method. Botanical origin did not have any major influence on the honey isotope profiles; only black locust honey showed higher δ(13)C values. Some differences were seen across different production years, indicating that the influence of season should be further tested. Statistical and multivariate analyses demonstrated differences among honeys of various geographical origins. Those from the Alpine region had low δ(13)C (-26.0‰) and δ(15)N values (1.1‰); those from the Mediterranean region, high δ(13)C (-24.6‰) and medium δ(15)N values (2.2‰); those from the Pannonian region, medium δ(13)C (-25.6‰) and high δ(15)N value (3.0‰); and those from the Dinaric region, medium δ(13)C (-25.7‰) and low δ(15)N values (1.4‰). 相似文献
10.
Maša Bošnjak Mojca Kržan Urša Lampreht Tratar Jožica Dolenc Maja Čemažar Alenka Seliškar 《Veterinary anaesthesia and analgesia》2021,48(1):35-41
ObjectiveTo investigate the pharmacokinetics of carprofen after a single intravenous (IV) dose and multiple oral doses administered to pigs undergoing electroporation of the pancreas.Study designProspective experimental study.AnimalsA group of eight female pigs weighing 31.74 ± 2.24 kg (mean ± standard deviation).MethodsCarprofen 4 mg kg?1 was administered IV after placement of a central venous catheter during general anaesthesia with isoflurane. Blood samples were collected 30 seconds before and 5, 10, 20, 30 and 60 minutes and 2, 4, 6, 8, 12 and 24 hours after carprofen administration. Subsequently, the same dose of carprofen was administered orally, daily, for 6 consecutive days and blood collected at 36, 48, 60, 72, 96, 120, 144 and 168 hours after initial carprofen administration. Plasma was analysed using liquid chromatography with mass spectrometry. Standard pharmacokinetic parameters were calculated by compartmental analysis of plasma concentration–time curves. Data are presented as mean ± standard error.ResultsThe initial plasma concentration of IV carprofen was estimated at 54.57 ± 3.92 μg mL?1 and decreased to 8.26 ± 1.07 μg mL?1 24 hours later. The plasma elimination curve showed a bi-exponential decline: a rapid distribution phase with a distribution half-life of 0.21 ± 0.03 hours and a slower elimination phase with an elimination half-life of 17.31 ± 3.78 hours. The calculated pharmacokinetic parameters were as follows: the area under the plasma concentration–time curve was 357.3 ± 16.73 μg mL?1 hour, volume of distribution was 0.28 ± 0.07 L kg?1 and plasma clearance rate was 0.19 ± 0.009 mL minute?1 kg?1. The plasma concentration of carprofen, administered orally from days 2 to 7, varied from 9.03 ± 1.87 to 11.49 ± 2.15 μg mL?1.Conclusions and clinical relevanceCarprofen can be regarded as a long-acting non-steroidal anti-inflammatory drug in pigs. 相似文献