Biochemical and physiological responses to long‐term Citrus tristeza virus infection in Mexican lime plants

Reactions that occur when a plant is subjected to Citrus tristeza virus (CTV) infection often result in triggering of numerous defence mechanisms to fight the infection. The reactions vary according to virus strain, host genotype, time of exposure to the infection and environmental conditions. To date, no study has examined in detail the consequences of 10‐year exposure to CTV infection on the biochemical and physiological status of susceptible Mexican lime plants (Citrus aurantifolia). To understand the reaction of such plants, changes in nutrient status, total proteins, enzyme activity involved in scavenging of reactive oxygen species, photosynthetic and transpiration rates, chlorophyll content, membrane permeability and water content were analysed in plants infected with different CTV isolates and in healthy plants. The activity of superoxide dismutase and polyphenol oxidase significantly decreased in the infected leaves, and membrane permeability was lower in the infected plants. Macro‐ and micronutrient elements were significantly changed: concentrations of leaf nitrogen, zinc, magnesium and iron were elevated but potassium concentration depressed in comparison to noninfected control leaves. Levels of other analysed nutrient elements, enzymes, photosynthesis and stomatal conductance, chlorophyll content and relative water content were unchanged. Clear physiological changes were found among infected and noninfected control plants but none between plants infected with different CTV isolates. The data suggest that some of the defence mechanisms investigated here were suppressed due to the continuous and long‐term pressure of biotic stress.     

The onset of puberty in Cameroon Dwarf goats kept as pets in northwestern Croatia

The aim of this study was to determine onset of puberty in Cameroon Dwarf goats (CDGs) kept as pets in northwestern Croatia by determining progesterone (PGS) and insulin‐like growth factor‐I (IGF‐I) levels in the blood by RIA methods. The first cyclic ovarian activity was estimated according to hormone profiles as determined in CDG in a moderate climate environment. Sixteen female CDG kids were kept in stables with access to pasture which provided space for exercise. The goat kids born in winter (December–January–February), spring (March–April–May), summer (June–July–August) and autumn (September–October–November) were assigned into four groups according to the season of the year of birth (n = 4 in each group). At 75 days of age at the initiation of the study, they weighed between 3.2 and 5.1 kg (4.24 ± 0.53 kg). The onset of ovulatory activity was determined by PGS and IGF‐I serum concentrations every 10 days starting from 75 days to 155 days of age. The onset of puberty in CDG kids occurred on average at 141.15 ± 2.66 days of age, but varied depending on the season of birth. All CDG kids born during summer were in heat by 155 days of age. In the other groups (kids born in autumn, winter or spring), one goat in each group was not in heat. Changes in blood serum PGS and IGF‐I concentrations during prepubertal and pubertal periods could aid in the evaluation of reproductive status and determination of the onset of puberty in CDG during all seasons of the year in a moderate climate region.     

Biological control of green mould on <Emphasis Type="Italic">Agaricus bisporus</Emphasis> by a native <Emphasis Type="Italic">Bacillus subtilis</Emphasis> strain from mushroom compost

Fifty bacterial isolates obtained from compost were tested in vitro against the causal agents of green mould in Agaricus bisporus. Isolate B-38 which induced 48.08% in vitro growth inhibition of T. harzianum T54 and 52.25% of T. aggressivum f. europaeum T77 was identified as Bacillus subtilis, based on 16S rDNA sequence and used in mushroom growing room experiments. B. subtilis B-38 did not decrease mycelial growth rate of Agaricus bisporus A15 in mushroom compost in glass tubes. After applying prochloraz-manganese, B. subtilis B-38 and B. subtilis QST 713, no significant differences in BE values among treatments were found concerning both total yield and the weight of healthy mushrooms. Statistical analyses showed that only inoculation significantly influenced the healthy mushroom yield. In plots inoculated with T. harzianum T54 disease incidence was significantly lower after treatments with prochloraz-manganese (11.81%), B. subtilis QST 713 (12.26%) and B. subtilis B-38 (14.19%) compared to the control (28.16%), as well as in plots inoculated with T. aggressivum f. europaeum T77 11.88%, 12.2% and 15.03%, respectively, in comparison with the control (23.47%). Statistically significant differences were not found among the efficacy values of tested bio-fungicides based on B. subtilis and the commercial fungicide prochloraz-manganese suggesting the use of B. subtilis B-38 and B. subtilis QST 713 as good alternatives to chemical fungicides.     

An Evaluation of the Effectiveness of Sorbents in the Remediation of Soil Contaminated with Zinc

Zinc exerts negative effects on soil and contributing to the degradation of soil ecosystems. New solutions for restoring healthy soil activity are therefore needed. The aim of this study was to evaluate the effectiveness of sorbents in the biological remediation of soil contaminated with zinc. A pot experiment was conducted on loamy sand. The tested plant was maize (Zea mays). Soil was contaminated with zinc chloride doses of 0, 100, 300, and 900 mg Zn²⁺ kg^?1 DM soil (dry matter of soil). Alginate, biochar, sepiolite, calcined halloysite, and a molecular sieve were added to soil in amounts corresponding to 2.5% of soil weight to minimize zinc’s potentially toxic effects on the biological properties of soil. The application of zinc stimulated the proliferation of all analyzed microbial groups. Zinc exerted negative effects on the ecophysiological diversity (EP) of fungi and the activity of dehydrogenases, catalase, and acid phosphatase. The applied sorbents modified the microbiological and biochemical properties of soil. In zinc-contaminated soil, alginate, biochar, and molecular sieve increased the counts of organotrophic, oligotrophic, and actinobacteria. Sorbents were not highly effective in promoting fungal growth and exerted varied effects on the activity of soil enzymes. The molecular sieve stimulated the activity of all soil enzymes, excluding β-glucosidase. Alginate minimized the negative influence of zinc on dehydrogenases and acid phosphatase, and biochar—on catalase, sepiolite, and calcined halloysite —on acid phosphatase. By modifying the biological properties of soil, the tested sorbents contributed to an increase in maize yields and a decrease in zinc uptake by maize plants.     

First report of invasive brown marmorated stink bug Halyomorpha halys (Stål, 1855) in Croatia

The brown marmorated stink bug, Halyomorpha halys (Stål, 1855) (Hemiptera: Heteroptera: Pentatomidae), has been spreading over Europe since the first documented records from Liechtenstein in 2004 and Switzerland in 2007. It is considered to be a pest in many agricultural crops and a household nuisance. In 2017 the species was observed in Croatia for the first time, in the city of Rijeka on the north Adriatic coast. Halyomorpha halys has already been recorded in three nearby countries (Italy, Hungary and Serbia), and therefore the arrival of this species had been expected from neighbouring populations or from distant sources via trading goods. To identify the pathway of entry, the haplotypes of H. halys (Stål, 1855) individuals were analysed by comparing a part of the mitochondrial COI gene with other haplotypes present in the GenBank database. Individual specimens shared the most common haplotype with nearby Italian and Hungarian populations.     

Viruses affecting tomato crops in Serbia

In a two-year survey (2011–2012), 3220 samples were collected and analyzed in order to determine the presence and distribution of viruses in tomato crops at 56 localities of 18 districts in Serbia. Out of 12 viruses tested, Cucumber mosaic virus (CMV), Potato virus Y (PVY), Alfalfa mosaic virus (AMV), Tomato spotted wilt virus (TSWV), Tomato mosaic virus (ToMV) and Tobacco mosaic virus (TMV) were detected in 42.1, 40, 11, 8.6, 2.3 and 1.3% of the total tested samples, respectively. The results revealed that CMV was prevalent in 2011 and PVY in 2012. CMV and PVY, apart from being predominant, were also the most widespread viruses. In general, single infections were the most frequent type of infection. Additionally, the most common mixed infections were double infections and the most prevalent combination was CMV and PVY. In 2011, the incidence of diseases and the percentage of all infection types were significantly higher than in 2012. Furthermore, in 2011, regardless of total single infections being prevalent compared to mixed infections, two prevailing viruses were commonly detected in mixed infections. The additional molecular testing of ELISA-negative samples using virus specific primers did not reveal the presence of Pepino mosaic virus (PepMV), Tomato yellow leaf curl virus (TYLC), Tomato infections chlorosis virus (TICV) and Tomato chlorosis virus (ToCV).     

Examining the Effects of the Destroying Ammunition,Mines and Explosive Devices on the Presence of Heavy Metals in Soil of Open Detonation Pit; Part 2: Determination of Heavy Metal Fractions

As a result of the destruction of ammunition, mines, and explosive devices by the method of open detonation, the increased concentration of heavy metals is often recorded in the soil of military polygons, which is a serious ecological problem. However, in order to determine the potential risk of such locations to the environment, it is necessary to determine, in addition to the total content, the forms in which the metals are present. In this paper, a sequential extraction method was used to analyze the six fractions of five heavy metals (cadmium, lead, nickel, copper, and zinc) in the soil of the polygon for destruction of ammunition, mines, and explosive devices. Samples were collected from the place of direct detonation (so-called pits) and from the edge of the pit. The aim of this research is determination of metal speciation in order to obtain a better insight in their mobility and risk arising from this. The results showed that heavy metals are predominantly present in the residual, oxide, and organic fractions. Cd and Cu were also significantly present in the mobile fractions due to conducted activities on the polygon. To assess the potential environmental risk of soil, the risk assessment code (RAC) and individual (ICF) and global (GCF) contamination factors were used. According to the RAC, the mobility and bioavailability of the analyzed heavy metals decreases in the following order: Cd?&gt;?Cu?&gt;?Zn?&gt;?Pb?&gt;?Ni. ICF results show low to moderate risk, while GCF results show low risk in terms of heavy metal contamination in the examined area.     

Effects of early leaf removal on grape yield,chemical characteristics,and antioxidant activity of grape variety Cabernet Sauvignon and wine from eastern Croatia

The aim of this study was to determine the impact of two different treatments of early defoliation performed before blooming on: grape yield, chemical parameters, polyphenols content, and antioxidant activity of grape and red wine cv. Cabernet Sauvignon from the vineyard located in Ilok, the eastern continental region of Croatia. Two different treatments of leaf removal (LR) were performed: removal of 3 leafs (T1) and 6 leafs (T2) before blooming, together with control (no leaf removal) (T3) during two years (2013 and 2014). Crop yield and average cluster weights per vine were determined. Density, pH and titratable acidity were measured in must, while the total phenols, total anthocyanins and antioxidant activity were measured in the extract of grape skin and produced wine. The analysis of individual anthocyanins in wine was performed by HPLC method. T2 treatment significantly lowered the crop yield and the average cluster weights, and increased total phenols, total anthocyanins, antioxidant activity and most abundant individual anthocyanins in wine. Defoliation did not affect the other chemical parameters in must, grape skin extract and wine. Vintage year is statistically the most significant source of variability for density of must, antioxidant activity in grape skin extract, as well as pH and titratable acidity in wine. This study has showed that the early leaf removal treatment in eastern continental part of Croatia could be used for the production of smaller quantity of high quality Cabernet Sauvignon red wine abundant with anthocyanins.