Molecular diversity of ‘Candidatus Phytoplasma pyri’ isolates in Slovenia

A European quarantine organism ‘Candidatus Phytoplasma pyri’ causing devastating pear decline disease has been reported to affect pear trees in several European countries. In this study a multilocus sequence analysis was successfully used to gain detailed insight into the molecular diversity of thirty closely related ‘Candidatus Phytoplasma pyri’ isolates from different orchards in Slovenia. Among three genomic regions analyzed, the 16S/23S rRNA intergenic spacer region was the most conserved among Slovenian isolates with 99.7 % sequence identity, yielding only three distinct genotypes. On the other hand, five different genotypes were detected when analyzing secY and aceF genomic regions that shared sequence identity of 94.8 and 97.2 %, respectively. Six of the detected genotypes, specifically four in the secY region and one in each of the two other analyzed genomic regions, were unique for Slovenia. At least eight different haplotypes were found with multilocus sequence analysis, indicating high molecular diversity among Slovenian ‘Ca. P. pyri’ isolates. Haplotypes were clustered into two major clusters, separated by at least 45 mutations. No connection was established between haplotype occurrence and cultivar type.     

Screening for antioxidant properties of Allium giganteum

The antioxidative properties of bulb, leaf and stalk of Allium giganteum were investigated. Activities of antioxidant enzymes (superoxide dismutase, catalase, peroxidase, glutathione peroxidase), quantities of malonyldialdehyde, superoxide and hydroxyl radicals and reduced glutathione and also the content of total flavonoids, chlorophylls a and b, carotenoids, vitamin C and soluble proteins were determined. The highest antioxidant activity was observed in the leaves. Furthermore, ESR signal of PBN-OH radical adducts in the presence of leaves phosphate buffer (pH 7) extract was reduced for 74.19%.     

Two outbreaks of neuropathogenic equine herpesvirus type 1 with breed-dependent clinical signs

Equine herpesvirus type 1 (EHV-1) is a worldwide spread pathogen of horses. It can cause abortion, respiratory and neurological disease and consequentially significant economic losses in equine industries. During 2009, two outbreaks of EHV-1 were confirmed in two stud farms in Eastern Croatia. The first outbreak occurred in February following the import of 12 horses from USA, serologically negative to EHV-1 before transport. Four mares aborted in the late stage of pregnancy and one perinatal death was recorded. Other six mares showed clinical signs of myeloencephalopathy with fatal end in four. One month later, the second EHV-1 outbreak was confirmed in stud farm about 100 km further with 17 abortions, three perinatal deaths and one mild neurological case. Epidemiological data showed that the disease was probably introduced in the first stud farm during international transport. The second outbreak started with the introduction of clinically healthy stallion from the first stud farm. Molecular characterisation and phylogenetic analysis confirmed that, despite different clinical signs, the identical virus caused both outbreaks. Both horse populations were free from EHV-1 infection before the outbreak and had not been vaccinated. Significant difference in clinical signs could be explained by different breed-related risk factors.     

Serum amyloid A isoforms in serum and milk from cows with Staphylococcus aureus subclinical mastitis

Serum amyloid A proteins (SAA) are very sensitive acute phase proteins, displaying multiple isoforms in plasma and different body fluids. They are currently under investigation as biomarkers of diseases. The aim of the present study was to compare the concentration and isoform expression of SAA in serum and milk of cows with bacteriologically negative milk (control group) and naturally occurring Staphylococcus aureus (S. aureus) subclinical mastitis (subclinical mastitis group). Somatic cell count (SCC) and bacteriological analyses were performed to establish the control and subclinical mastitis group. SAA concentration was evaluated using a commercial ELISA kit, while expression of different isoforms (serum A-SAA and milk M-SAA3 isoforms) was visualized by denaturing isoelectrical focusing and immunoblotting. The SAA concentrations in sera and milk of cows in the subclinical mastitis group were three and 100 times higher than in those from the control group of cows, respectively. Cows in the subclinical mastitis group had more acidic SAA isoforms in serum with the most prominent one at pI 5.5. This isoform was not detected in sera from the control group. Milk samples in the subclinical mastitis group contained abundant highly alkaline M-SAA3 isoforms and most of the serum isoforms, except for that at pI 5.5. In the subclinical mastitis group SAA isoforms with equivalent pI as serum isoforms accounted for 20% of the total SAA concentration in milk. There were significant differences in the concentrations and isoform patterns of SAA in serum and milk between the control and subclinical mastitis groups of cows. Also, we demonstrated that serum SAA isoforms were not transferred to milk proportion to their plasma content.     

Effects of alternative pesticides on greenhouse whitefly in protected cultivation

The greenhouse trials were conducted to determine the efficacy of environmentally safe insecticides/fungicides with physical mode of action [Agri-50E (propylene glycol alginate, hydrated) and SB Plant Invigorator] against Trialeurodes vaporariorum (Westwood). The efficacy of Agri-50E on adults and nymphs (first, second, third, and fourth instars) was compared to the commercially available standard Boxer 200 SL (imidacloprid) and an untreated control on greenhouse-grown poinsettia. Insecticides were applied by foliar spraying, Agri-50E (2 or 3 ml/l) two times and Boxer 200 SL (1 ml/l) once. Three and 7 days after first application (DAFA), imidacloprid was more effective than both concentrations of Agri-50E in controlling T. vaporariorum adults, whereas 10 DAFA there were no differences between insecticides. Twelve DAFA, imidacloprid was more effective than Agri-50E in controlling first, second and third instars and after 19 days in controlling second, third, and fourth instars. The results obtained from the poinsettia study, indicated that Agri-50E (2 or 3 ml/l) is successful in controlling T. vaporariorum adults and moderately effective against T. vaporariorum immature stages. In the second experiment, the efficacy of Agri-50E on adults was compared to SB Plant Invigorator (SBPI), the standard treatment Calypso 480 SC (thiacloprid), and an untreated control on greenhouse-grown tomato. Agri-50E (3 ml/l) and SBPI (2 ml/l) were applied four times while Calypso 480 SC (0.4 ml/l) was applied twice. The results obtained from tomato study showed that SBPI was the most effective insecticide against T. vaporariorum adults, followed by Agri-50E and Calypso 480 EC. Beside the ecological benefits, the alternative insecticides Agri-50E and SBPI might have an important role to play in future pest management programs.     

Relationships between line per se and testcross performance for agronomic traits in two broad-based populations of maize

Degree of the association between line per se performance (LP) and testcross performance (TP) is important in breeding programs and simultaneous improvement of commercial hybrids and their parental lines. This experiment was designed to study genetic variability and genetic correlation for several agronomic traits in two maize (Zea mays L.) broad-based populations (NS12-SG and NS14-SG). Independent trials with 80 entries of S₁ progenies as well as their testcrosses were arranged according to an incomplete block design with replicates in sets. Grain yield, stay green, anthesis-silking interval, stalk water content and grain moisture were evaluated in four environments. The anthesis-silking interval had the highest genetic variation, followed by stay green. High heritability estimates (>0.50) for all traits, pointed out that further selection would be possible. Genetic correlations between line per se and testcross performance were lowest for grain yield (0.396** and 0.592**, for NS12-SG and NS14-SG, respectively), and highest for grain moisture (0.937** and 0.821**, respectively). High correlations between line per se and their testcrosses for stay green, anthesis-silking interval, stalk water content and grain moisture indicated that additive gene action might be more important than dominance in controlling the expression of these traits.     

Fungicide sensitivity,growth rate,aggressiveness and frost hardiness of <Emphasis Type="Italic">Monilinia fructicola</Emphasis> and <Emphasis Type="Italic">Monilinia laxa</Emphasis> isolates

Monilinia fructicola, the most destructive pathogen of the genus Monilinia, has recently been introduced into Serbia and many other European countries. Since then, many studies have been conducted to evaluate the characteristics of Monilinia species that have a role in the establishment and survival of the pathogen in new areas. The present study assessed the capacity of M. fructicola to repress and replace Monilinia laxa in Serbia based on: fungicide sensitivity, growth rate and aggressiveness at different temperatures, as well as frost hardiness of the isolates of both species. The results showed that the isolates of M. fructicola, compared to M. laxa, were significantly less sensitive to the following fungicides: iprodione, tebucanozole, chlorothalonil, azoxystrobin, fluopyram, and boscalid. In addition, M. laxa isolates exhibited little variation in sensitivity to all of the tested fungicides, whereas M. fructicola isolates displayed a wide range of sensitivity. The temperature of 5°C favored M. laxa growth and aggressiveness, while at 30°C M. fructicola grew faster and had higher lesion expansion rate. These results support an assumption that M. fructicola will continue to spread in Serbian orchards in coming years, particularly on stone fruits harvested during hot summer weather.     

Examining the Effects of the Destroying Ammunition,Mines, and Explosive Devices on the Presence of Heavy Metals in Soil of Open Detonation Pit: Part 1—Pseudo-total Concentration

This paper presents the results of determining the pseudo-total concentration of five heavy metals in the soil on which the destruction of ammunition, mines, and explosive devices is carried out by the method of open detonation. In the analyzed area, the concentrations of cadmium, lead, nickel, copper, and zinc were determined, while from the physical properties of the soil were determined the granulometric composition and the pH. The aim of the study is to determine the origin and total load on heavy metals and, based on that, to assess the dangers and impact of the site in terms of the soil pollution by heavy metals. In accordance with the regulations of Bosnia and Herzegovina, the results of the soil testing showed a significant load of copper (up to seven times) and cadmium (up to six times), and exceeding the allowed values for nickel and zinc in some places. Lead was the only metal whose concentration was within the maximum allowed and according to that the soil was classified as unpolluted. A sample of soil from the edge of the pit is the only sample in which all heavy metals, except Ni, were within the maximum allowable concentration. In regard to the concentration of the examined metals, the soil of the pit is classified as medium polluted from the aspect of copper, cadmium, and nickel and highly contaminated with zinc. The concentrations of copper and zinc in the examination area correspond to contaminated soil that represents ecological risk, which requires soil remediation.     

Plant Accumulation of Natural Radionuclides as Affected by Substrate Contaminated with Uranium-Mill Tailings

Environmental concern due to plant accumulation of natural radionuclides is a major concern in uranium mining areas. To evaluate the risk associated with the transfer of radionuclides to edible plants, the uptake of ²³⁸U, ²²⁶Ra, and ²¹⁰Pb by Chinese cabbage (Brassica rapa L. subsp. pekinensis (Lour.) Hanelt) grown in soils contaminated with uranium-mill tailings (UMT) was investigated. Test plants were grown under controlled conditions in substrate composed of soil and UMT in different ratios. Activity concentrations of ²³⁸U, ²²⁶Ra, and ²¹⁰Pb in substrate, leaves, and roots were measured and the concentration ratios determined. Soil characteristics were determined, since they directly affect bioavailability of radionuclides. Concentration ratios of ²³⁸U, ²²⁶Ra, and ²¹⁰Pb in leaves varied from 0.001 to 0.006, 0.024 to 0.172, and 0.004 to 0.011, respectively, and in roots from 0.020 to 0.126, 0.015 to 0.241, and 0.033 to 1.460, respectively. Concentrations of ²³⁸U, ²²⁶Ra, and ²¹⁰Pb in leaves and roots were found to correlate with the amount of ²³⁸U, ²²⁶Ra, and ²¹⁰Pb in the substrate. A higher amount of ²²⁶Ra accumulated in aboveground parts (57–877 Bq kg^?1 d. m. for leaves) compared to ²³⁸U (0.6–4.7 Bq kg^?1 d. m. for leaves) and ²¹⁰Pb (8–53 Bq kg^?1 d. m. for leaves), which were mainly stored in the roots. The relationships between the amount of radionuclides in plants and soil characteristics and their role in radionuclide uptake are discussed and critically evaluated.