Chestnut blight fungus in Croatia: diversity of vegetative compatibility types,mating types and genetic variability of associated Cryphonectria hypovirus 1

In order to improve understanding of its diversity, 338 isolates of Cryphonectria parasitica, the causal agent of chestnut blight, were sampled from 10 chestnut populations throughout chestnut‐growing coastal and continental areas of Croatia. Eighteen vegetative compatibility (VC) types were identified. The VC type EU‐1 was the most widespread, comprising 42·9% of the isolates, followed by EU‐2 (21%) and EU‐12 (14·2%). In respect to the occurrence of the main VC types, the C. parasitica populations in Croatia combined features of both northwestern and southeastern European populations. Perithecia and mating‐type ratios of approximately 1 : 1 were found in all populations, suggesting that sexual reproduction of the fungus is common in Croatia. Natural hypovirulence was also evident in all populations, with incidence of hypovirus‐infected isolates ranging from 12·7% in Istria‐Buje to 66·6% in the continental part of the country. A total of 36 hypovirus‐infected isolates sampled throughout Croatia were analysed in ORF‐A and ORF‐B by RT‐PCR/RFLP analysis. All viral isolates belonged to the Italian subtype of Cryphonectria hypovirus 1 (CHV‐1) and were closely related to the isolates found in other European countries. The RFLP patterns found were also identical or similar to the patterns of three isolates collected in Croatia 22 years ago, suggesting a slow evolution of the hypovirus.     

Effect on neonatal oestrogen and hCG treatment on the genital organs of adult female rats

The ovaries and the uterine as well as vaginal mucous membranes of 80-, 180- and 365-day-old intact female rats and females neonatally treated with a single dose of oestrogen and repeated doses of human chorionic gonadotropin (hCG) were studied. Numerous follicles, interstitial cells and corpora lutea (CL) were present on the ovary of intact females from 80 up to 365 days of age. The number of primary and secondary follicles decreased in the intact female rats between Day 180 and Day 365 of life. On the ovaries of 180- and 365-day-old female rats neonatally treated with oestrogen, interstitial glandular cells and cystic follicles predominated. No CL were present on these ovaries. The height of epithelial cells of the uterine and vaginal mucous membranes increased in intact female rats from 80 to 365 days of age, whereas in oestrogen-treated females the height of epithelium decreased. From 80 up to 365 days of age, the height of epithelial cells of uterine and vaginal mucous membranes of rats neonatally treated with repeated doses of hCG was similar to that in the corresponding control animals.     

The study of coloration and antibacterial efficiency of corona activated dyed polyamide and polyester fabrics loaded with Ag nanoparticles

The aim of this study was to examine the influence of dyeing on antibacterial efficiency of corona activated polyamide and polyester fabrics loaded with colloidal Ag nanoparticles as well as the influence of the presence of Ag nanoparticles on the color change of dyed fabrics. C.I. Acid Green 25 and C.I. Disperse Blue 3 were used for dyeing of polyamide fabrics and C.I. disperse violet 8 for polyester fabrics. The color change of polyamide fabrics depends on the dye type, which was generally lower compared to polyester fabrics. Antibacterial efficiency of Ag loaded fabrics was tested against Gram-positive bacterium Staphylococcus aureus and Gram-negative bacterium Escherichia coli. Corona activated polyester and polyamide fabrics showed excellent antibacterial efficiency independently of order of dyeing and Ag loading. The morphology of fibers loaded with Ag nanoparticles was assessed by SEM and atomic absorption spectroscopy for elemental analysis.     

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.     

Dependence of the common carp (Cyprinus carpio L.) fatty acid profile on diet composition in a semi‐intensive farming system: tissue and time variability

Increasing the amount of n‐3 PUFA in carp meat is one of the most important tasks in enhancing its overall quality. However, the relative influence of supplemental feed and natural feed on the FA profile of cultivated carp flesh is not well documented, making it difficult to choose the most efficient strategy. Carp diet composition and diet and flesh fatty acid profiles were simultaneously followed in order to determine the influence of pelleted feed compared to natural feed on fatty acid profiles of carp tissues. Pelleted feed clearly dominated over zooplankton and Chironomidae in the carp diet, producing carp tissue fatty acid profiles closely resembling those of pelleted feed. However, increase in the abundance of zooplankton over the course of the investigation resulting in concomitant increase of its proportion in feed bulk was in strong positive correlation with increase in n‐3 HUFA in the dorsal muscle. n‐3 PUFA enrichment of supplemental feed could be the dominant mechanism in production of carp meat rich in n‐3 PUFA. However, even a small increase in natural feed availability could significantly change the diet of common carp and result in considerable improvement of meat quality with regard to n‐3 HUFA content.     

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.     

Effects of fish farming on microbial planktonic communities in the middle Adriatic sea

The effects of farming on planktonic microbial communities were investigated at the coastal sea bass/sea bream farm in the oligotrophic middle Adriatic Sea. Analyses of nutrients, chlorophyll a, cyanobacteria (Prochlorococcus), heterotrophic bacteria and nanoflagellates (both pigmented and heterotrophic) were carried out on samples taken during six seasonal cruises at different water depths. Farming activity increased the natural concentrations of some nutrients (ammonium, nitrite, phosphate, dissolved inorganic nitrogen) and changed their seasonal pattern. The values still remained typical for oligotrophic environ‐ments demonstrating no risk for eutrophication. Enhanced nutrient supply provoked an immediate increase in abundances of both autotrophic and heterotrophic microbial groups (except pigmented nanoflagellates) and chlorophyll a. The effect of farming was more visible for the heterotrophic microbial component. Results from this study reveal a potential common pattern of microbial response to farming in the oligotrophic environments. It also suggests the importance of heterotrophic microbial web in transferring the matter and energy released from the fish farms in nutrient poor environments.     

Using trace elements in otoliths to discriminate between wild and farmed European sea bass (<Emphasis Type="Italic">Dicentrarchus labrax</Emphasis> L.) and Gilthead sea bream (<Emphasis Type="Italic">Sparus aurata</Emphasis> L.)

Trace elements in otoliths of sea bass (Dicentrarchus labrax L.) and sea bream (Sparus aurata L.) from fish farms and coastal wild populations in the western Mediterranean Sea were analysed by inductively coupled plasma-mass spectrometry. Results showed that concentrations of Mg, K, and Mn differed significantly between wild and farmed sea bass, while concentrations of Mg, K, Mn, Fe, Zn, Sr, and Ba varied significantly between wild and farmed sea bream. Discriminate analysis and cross-validation classification showed that the trace element profile in otoliths can be used to separate farmed fish from wild stocks with high accuracy on both sea bass (individuals correctly classified: 90.7 %) and sea bream (individuals correctly classified: 96.6 %). Moreover, trace elements in otoliths resulted to be useful to discriminate among wild fish stocks within each species.     

Distribution of porcine circovirus 2 cap antigen in the lymphoid tissue of pigs affected by postweaning multisystemic wasting syndrome

The lymphatic organs of 50 pigs from a total of eight farms located at different sites in the epizootiological region of North Ba?ka County were studied to obtain data on the prevalence of circoviral infections in Serbia. All of the pigs examined had clinical signs suggestive of postweaning multisystemic wasting syndrome (PMWS). All pigs underwent necropsy and tissue samples were taken for histopathological, immunohistochemical (IHC) and PCR analysis. The presence of porcine circovirus 2 (PCV2) was established by PCR analysis in the organs of the pigs tested. The most frequent histopathological lesions of lymphoid tissue linked with the presence of positive immunostaining for PCV2 Cap antigen confirmed the existence of PMWS in all farms tested in North Ba?ka County. Using PCR, histopathological and IHC techniques, the presence of PMWS was proved in the Republic of Serbia. During necropsy, generalised enlargement of the lymph nodes was evident. The most common histopathological finding was lymphocyte depletion in the follicular and perifollicular areas of lymph nodes. Infiltration by macrophages was also recorded. By IHC analysis, the cytoplasm of macrophages was shown to contain a large amount of the ORF2-coded Cap antigen of PCV2. Lymphocyte depletion and large numbers of macrophages were recorded in the tonsils, spleen, intestinal lymphatic tissue, Peyer's patches and ileocaecal valve. The presence of typical granulomatous lesions with multinuclear giant cells (MGCs) was also recorded in the lymphatic tissue. Cap antigen was shown to be present in macrophages and less often in lymphocytes.     

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.