Ágnes Farkas

Detection of non-polio enteroviruses in Hungary 2000–2008 and molecular epidemiology of enterovirus 71, coxsackievirus A16, and echovirus 30

Human enteroviruses are associated with various clinical syndromes from minor febrile illness to severe, potentially fatal conditions like aseptic meningitis, paralysis, myocarditis, and neonatal enteroviral sepsis. Between June 2000 and August 2008 echovirus (E) type 2, 4, 6, 7, 9, 11, 13, 25, 30, coxsackievirus (CV) -A16, -A19, -B5, and enterovirus 71 (EV71) were reported in Hungary. In this study, 29 previously enterovirus positive samples from 28 patients diagnosed with hand, foot and mouth disease, meningitis and encephalitis, were molecularly typed. The genetic relationships of identified serotypes CV-A16, EV71, and E30 were assessed by direct sequencing of genomic region encoding the capsid protein VP1. The sequences were compared to each other and sequences from other geographical regions possessed in Genbank. The phylogenetic analysis of CV-A16 revealed that the viruses were mostly of Far-Eastern or Asia-Pacific origin. Typing of EV71 showed that one virus from 2000 belonged to genotype C1 and five viruses observed in 2004 and 2005 were identified as genotype C4. The 11 echovirus 30 strains showed homology with those of neighbor European countries. The molecular examination of E30 revealed that three separate lineages circulated in 2000, 2001, and 2004–2006 in Hungary.

Determination of tropane alkaloids atropine and scopolamine by liquid chromatography-mass spectrometry in plant organs of Datura species

Hyoscyamine (atropine) and scopolamine are the predominant tropane alkaloids in the Datura genus, occurring in all plant organs. The assessment of the alkaloid content of various plant parts is essential from the viewpoint of medical use, but also as a potential risk of toxicity for humans and animals. Therefore, a reliable method for the determination of tropane alkaloid content is of high importance. The present work aimed at the elaboration of a rapid method for determination of the most abundant Datura alkaloids by LC-MS technique using a new generation of core-shell particle packed column. Tropane alkaloid content was investigated in various plant organs of four Datura taxa (D. innoxia, D. metel, D. stramonium, and D. stramonium var. tatula), grown under the same conditions, in two developmental stages. We have developed a rapid LC-MS method for the quantitative determination of atropine and scopolamine, which was successfully applied to quantify the alkaloids in different plant organs (leaves, flowers, stems, seeds) of thorn apples after a simple sample preparation step. Elaboration and validation of the method and analysis of plant extracts were done by UFLC-MS technique, employing an Ascentis Express C18 column. Detection was done in positive ionization mode (ESI+) and the method suitability was evaluated by several validation characteristics. Quantitation limits are 333 and 167 pgmL(-1) for scopolamine and atropine, respectively, and the method shows very good repeatability. The analysis of Datura extracts revealed significant differences depending on the species, the organ and the sampling period. Atropine was found to be dominant over scopolamine in three out of the four taxa investigated. D. innoxia showed the highest concentrations of scopolamine in all organs examined, whereas D. metel accumulated the lowest scopolamine levels. Hyoscyamine, measured as atropine, was the highest in D. stramonium var. tatula, and the lowest in D. innoxia. Samples collected in summer had higher scopolamine levels than autumn samples, concerning both stems and leaves.

Nectar secretion dynamics of Hungarian local pear cultivars

Abstract. The dynamics of daily nectar secretion was studied in 28 local pear cultivars in a Hungarian cultivar collection and three main types were distinguished. 1. Cultivars secreting nectar continuously are the most favourable for pollinators, rewarding them both with nectar and pollen. 2. Cultivars with discontinuous nectar secretion have sufficient insect attraction only if anther dehiscence is continuous during the day. In the first two types nectar secretion peaks usually appeared at 9:00 or 10:00, 14:00 or 15:00 and 18:00 or 19:00, with 4–5-hour intervals. 3. In some cultivars no nectar secretion was observed in any or some of the studied years, these being the least attractive for pollinators. The cultivars studied did not necessarily show the same type of nectar secretion dynamics in each season. From the viewpoint of pollination it is also of high importance, in which developmental phase nectar secretion begins.

An ethnobotanical study on home gardens in a Transylvanian Hungarian Csángó village (Romania)

Due to their isolation and insufficiency of official medical care, the inhabitants of several Transylvanian “Csángó” farms in Romania have valuable archaic knowledge of plants. Such ethnobotanical data reported previously from different regions of the country are available only in Hungarian. Of the various ethnic groups constituting the Csángós’ culture the present study was undertaken to survey those living in the Úz-valley (Romania), focusing on the indigenous human ethnomedicine and ethnoveterinary practices of Csinód, a village in the Eastern Carpathian Mountains. The aim of the study was to summarize the occurrence and diversity of food, medicinal, ornamental and fodder plants, as well as herbal home remedies applied on a daily basis. In the summers of 2007–2009 altogether 85 plant taxa in 13 home gardens were registered, using free interviews with local terminology, concentrating on drug parts, use, origin of knowledge and peculiar magico-mythological procedures associated with the species concerned. Due to the obvious decrease in the villagers’ traditional knowledge the prevention of these data from disappearing has gained primary importance in an effort to preserve this heritage as an integral part of the folk medicinal system in Transylvania.

Surveillance of human rotaviruses in 2007–2011, Hungary: Exploring the genetic relatedness between vaccine and field strains

BACKGROUND The availability of rotavirus vaccines has resulted in an intensification of post vaccine strain surveillance efforts worldwide to gain information on the impact of vaccines on prevalence of circulating rotavirus strains. OBJECTIVES In this study, the distribution of human rotavirus G and P types in Hungary is reported. In addition, the VP4 and VP7 genes of G1P[8] strains were sequenced to monitor if vaccine-derived strains were introduced and/or some strains/lineages were selected against. STUDY DESIGN The study was conducted in 8 geographic areas of Hungary between 2007 and 2011. Rotavirus positive stool samples were collected from diarrheic patients mostly <5 years of age. Viral RNA was amplified by multiplex genotyping RT-PCR assay, targeting the medically most important G and P types. When needed, sequencing of the VP7 and VP4 genes was performed. RESULTS In total, 2380 strains were genotyped. During the 5-year surveillance we observed the dominating prevalence of genotype G1P[8] (44.87%) strains, followed by G4P[8] (23.4%), G2P[4] (14.75%) and G9P[8] (6.81%) genotypes. Uncommon strains were identified in a low percentage of samples (4.12%). Phylogenetic analysis of 318 G1P[8] strains identified 55 strains similar to the Rotarix strain (nt sequence identities; VP7, up to 97.9%; VP4, up to 98.5%) although their vaccine origin was unlikely. CONCLUSIONS Current vaccines would have protected against the majority of identified rotavirus genotypes. A better understanding of the potential long-term effect of vaccine use on epidemiology and evolutionary dynamics of co-circulating wild type strains requires continuous strain surveillance.

Detection of Human Bocavirus from Fecal Samples of Hungarian Children with Acute Gastroenteritis

Objectives: Human bocavirus (HBoV), a newly identified member of the Parvoviridae family is associated with respiratory tract and gastroenteric infections, mostly of young children. HBoV infections show a seasonal distribution with the peak in temperate areas being in the winter months. Methods: In our study, 35 throat swabs from children under 5 years with acute respiratory symptoms and 61 stool samples from children (<5 years) with acute gastroenteritis were collected in the period of October 2007–March 2008. A HBoV-specific polymerase chain reaction for detection of the virus, and sequence analysis for identification of virus variants were performed. Results: Although respiratory samples were all negative, 3.3% of stool samples (2/61) proved to be positive for HBoV. The virus carrier children were 3 and 5 years old. The ratio of HBoV positive samples is similar to international results (2.1–5.5%). Conclusions: According to the result of sequence analysis of HBoV, the occurrence of genotype 2 of HBoV in Hungary is confirmed.

Floral traits affecting fire blight infection and management

Erwinia amylovora, the causative agent of fire blight, colonizes primarily the flowers of the sub-family Maloideae. Commercially important fruit tree species such as apple (Malus domestica) and pear (Pyrus communis) are also affected by the disease. Epiphytic bacterial populations develop on the stigma, from where the pathogen colonizes the hypanthium, aided by moisture. Under favorable conditions, nectar provides a rich medium for growth, which allows bacterial invasion of tissues through the stomata of the nectary. The paper reviews various floral traits that may play a role in the onset and progression of the infection. Flower age, stigma morphology and longevity, the size of epiphytic bacterial population, morphology of the hypanthium, anatomy of the nectary, dynamics of nectar secretion, as well as the volume, concentration and composition of the nectar are discussed in detail, comparing traits of susceptible versus tolerant apple and pear cultivars. Management programs, aiming at the suppression of E. amylovora on floral parts by antibiotics, chemical compounds, natural substances or biological control agents, are also discussed.

Variation in Nectar Volume and Sugar Concentration of Allium ursinum L. ssp. ucrainicum in Three Habitats

Floral nectar volume and concentration of ramson (Allium ursinum L. ssp. ucrainicum) were investigated in three different habitats, including two types of sessile oak-hornbeam association on brown forest soil with clay illuviation and a silver lime-flowering ash rock forest association on rendzina. Daily nectar production ranged from 0.1 to 3.8 μL per flower with sugar concentrations of 25 to 50%. Mean nectar volumes and concentrations showed significant differences between freely exposed flowers and covered flowers, which had been isolated from flower visitors 24 h prior to nectar studies. Both the amount and quality of nectar were affected by microclimatic conditions and soil properties and varied between populations at different habitats. In the silver lime-flowering ash rock-forest association mean nectar volumes and concentrations were lower than in a typical sessile oak-hornbeam association on three occasions, the difference being significant in two cases. During full bloom, the date of sampling did not have a profound effect on either nectar volume or concentration.

Tall Wheatgrass Cultivar Szarvasi-1 (Elymus elongatus subsp. ponticus cv. Szarvasi-1) as a Potential Energy Crop for Semi-Arid Lands of Eastern Europe

By 2020, proportion of renewable energy sources should be around 20 per cent of the total energy consumption in the European Union, according to the new treaty signed by European leaders in 2009. This vast amount of renewable energy can be sourced from hydroelectric, geothermal, wind, solar power and, of course, from biofuels. To achieve this ambitious target, new technologies must be invented to exploit energy from the abiotic source of renewables and new energy plant species should be developed and produced, serving as source for solid, liquid biofuels and for biogas production. The most intensively studied and used bioenergy crops include miscanthus, reed canary grass, willows and poplars. We already have considerable knowledge about these energy plants from their taxonomical relations to their detailed crop technologies. In this chapter, we introduce a novel energy plant that has been cultivated for more than a century in many parts of the world for numerous purposes (e.g. land remediation, erosion control, forage), but its potential for energy production has not yet been realized. Tall wheatgrass, a new energy crop (Elymus elongatus subsp. ponticus cv. Szarvasi-1) has recently been introduced to cultivation in Hungary to provide biomass for solid biofuel energy production. The cultivar was developed in Hungary. The main goal of our research was to investigate the performance of Szarvasi-1 energy grass under different growing conditions (e.g. soil types, nutrition supply). We focused on the ecological background, biomass yield, weed composition, morphology, ecophysiology and the genetics of the plant.

Detection and sequencing of West Nile virus RNA from human urine and serum samples during the 2014 seasonal period

West Nile virus, a widely distributed mosquito-borne flavivirus, is responsible for numerous animal and human infections in Europe, Africa and the Americas. In Hungary, the average number of human infections falls between 10 and 20 cases each year. The severity of clinically manifesting infections varies widely from the milder form of West Nile fever to West Nile neuroinvasive disease (WNND). In routine laboratory diagnosis of human West Nile virus infections, serological methods are mainly applied due to the limited duration of viremia. However, recent studies suggest that detection of West Nile virus RNA in urine samples may be useful as a molecular diagnostic test for these infections. The Hungarian National Reference Laboratory for Viral Zoonoses serologically confirmed eleven acute human infections during the 2014 seasonal period. In three patients with neurological symptoms, viral RNA was detected from both urine and serum specimens, albeit for a longer period and in higher copy numbers with urine. Phylogenetic analysis of the NS3 genomic region of three strains and the complete genome of one selected strain demonstrated that all three patients had lineage-2 West Nile virus infections. Our findings reaffirm the utility of viral RNA detection in urine as a molecular diagnostic procedure for diagnosis of West Nile virus infections.