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The molecular background of depression is intensively studied in terms of alterations of inhibitory circuits, mediated by gamma aminobutyric acid (GABA) signalization. We investigated the effects of chronic social isolation (CSIS) and chronic fluoxetine (Flx) treatment (15 mg/kg/day) (3 weeks), on Parvalbumin (PV) and GAD67 expression in a layer-specific manner in rat dorsal hippocampal subregions. CSIS-induced depressive- and anxiety-like behaviours were confirmed with decrease in sucrose preference and increase in marble burying during behavioural testing, while Flx antagonized these effects. CSIS altered PV expression in stratum pyramidale (SP) of dorsal cornu ammonis 1 (dCA1) and stratum radiatum (SR) of dCA3. Flx antagonized this effect, and boosted PV expression in SP of the entire dCA and the dorsal dentate gyrus (dDG), as well as in the SR of dCA1/CA3. CSIS showed no significant effects on GAD67 expression, while Flx boosted its expression within the SR of the entire CA and SO of the dCA3. A correlation between SP of dCA1 and SR of dCA3 with regard to PV changes, implicates their possible role in the inhibitory circuit alterations. Flx-induced increase in GAD67 expression, specifically in SR of the entire dHIPP, may impose its involvement in the cell metabolic processes. Strong negative correlation between GAD67 and sucrose preference following Flx-treatment of CSIS rats was revealed. PV + cells of the SP layer of dCA1 and CA2 could be a potential target for the antidepressant action of Flx, while strong effect of Flx on GAD67 expression in the SR should be more extensively studied.

This research aimed to test whether coating with Lactobacillus paracasei subsp. paracasei BGHN14 homogenate may increase soluble protein and amino acid amount on larval fish feed surface. Total amino acid and protein, as well as taste stimulating and growth promoting amino acid amounts were analyzed in coated feed samples. Results indicated that coating with BGHN14 homogenate increased the amount of soluble proteins and free Glycine on feed surface, as well as the availability of protein-bound (hydroxy-)Proline. This lab-scale research provides the basis for use of lactobacilli as resource efficient source of soluble nutrients for animal feedstuffs.

Within the nursery sector, a global interest in functional fruits imposes a permanent need to improve the tissue culture technology. A modern bioreactor immersion system is currently a valid solution for the micropropagation of many cultures. At the Faculty of Agrobiotechnical Sciences Osijek (FAZOS, Croatia), a research was conducted with the aim of improving the raspberry (Rubus idaeus L., cv. HimboTop®) micropropagation protocol. The treatments implied the modifications of the MS and DKW nutrient medium (DKW – standard; DKWm – substitution of FeEDTA with the FeEDDHA; MSm – macroelement modification, 3 x CaCl2, MgSO4, and KH2PO4 + FeEDDHA), differently depending on the aggregate condition (semi‐solid and liquid SETISTM). Subsequent to 15 (SETISTM) and 30 cultivation days (semi-solid medium), respectively, a morphological parameter analysis and a multiplication success evaluation were commenced. In all treatments (DKW, DKWm, and MSm), the observed parameters (shoot height, number of leaves, leaf width and length) were significantly better in an interaction with the liquid medium. Comparing the models, a liquid medium (SETISTM) resulted in a significantly higher explant multiplication (4.02) (a semi‐solid medium - 2.74). The best reproduction quality, implying a better plant vigor with a reduced vitrification, was obtained on the MSm medium in the SETISTM. A concentration optimization of individual mesocomponents (i.e., of the macroelements) and their chemical forms (FeEDDHA) may contribute to their improved adoption and consequently to the stress suppression due to explant transplantation. Selecting the modern technical solutions (TIS bioreactors - SETISTM) and continuously improving the protocols, we may obtain the realistically sustainable models, suitable for a massive clonal reproduction.

The aim of this study was to determine the influence of the origin and size of planting material on the morphological characteristics of seed tuber's three potato cultivars. The objects of research were three varieties of potatoes: Aladdin, Newton and Desiree. The analysis of the morphological characteristics tuber showed highly significant (r

Fire blight disease, caused by plant pathogenic bacteria Erwinia amylovora represents a global threat for pome fruit industry, affecting yield and causing serious economic damage. During six year period (2011–2016), a total of 45 isolates typical for E. amylovora were isolated from four different host plants (apple, pear, quince and medlar) in 21 localities within four Serbian regions (Southeast, Central, West, and North). Isolates were primarily identified using biochemical and serological tests, polymerase chain reaction (PCR) with species-specific primer pair (A/B) and their pathogenicity. Polyclonal antibodies used in DAS ELISA and IF were found to be specific and sensitive enough for rapid E. amylovora detection. In virulence test performed on immature pear fruitlets, higher variability (CV) was found between regions than among hosts from which isolates originated. Slighter virulence of isolates originated from North region of Serbia (Vojvodina) was found in comparison to the isolates from Central Serbia, which were the most variable. Principal components analysis (PCA) segregated quince as a host among the most prevalent isolates in group of highly virulent, but failed to identify difference related to geographic origin of Serbian isolates. The same results enabled multivariate statistical analysis. Different DNA fingerprinting techniques such as repetitive element sequence based PCR (rep-) with BOX, ERIC, REP, GTG5 and SERE primers, then Random Amplified Polymorphic DNA (RAPD) with M13-PCR primer revealed genetic homogeneity among all Serbian E. amylovora isolates. ERIC, SERE and M13 primers generated the most complex and visually perceptible patters. Restriction fragment length polymorphism analysis (RFLP) of gyrA and recA genes, digested with three restriction endonucleases BstUI, BgIII and BsmI and rpoS gene digested with BstUI gave the unuque, characteristic patterns for all Serbian isolates.

Tests of seed germination, seed dormancy and seedling growth were performed on 0-, 6-, 20- and 30-months-old seed lots of Festulolium in comparison with Italian ryegrass (Lolium multiflorum) and meadow fescue (Festuca pratensis). Tests were performed on seeds harvested in 2 different years (2014 and 2015) resulting in no major difference between the years. Seed storage affected seed viability and dormancy and seedling growth in all 3 grasses. The maximum germination of Festulolium seeds was achieved 6 months after harvest (95% normal seedlings); germination decreased significantly thereafter. While maximum germination of L. multiflorum and F. pratensis seeds was also achieved following storage for 6 months, these germination rates (93 and 90%, respectively) were retained until at least 20 months in storage. After storage for 30 months, seed germination of Festulolium, L. multiflorum and F. pratensis had declined to 72, 79 and 83%, respectively. High germination in all species was associated with higher rates of seedling growth. In an artificial seed ageing test, a temperature of 41 °C (during 48 and 72 hours) was found to effectively rank seed lots for germination performance in all 3 grasses. This test seems to have application for use in the seed trade to identify seed lots which could deteriorate more rapidly in storage. Further studies are needed to verify this hypothesis.

The objective was to assess the effect of chemical desiccation on seed germination and storability in three sunflower female inbred lines and to point at possible indicators for optimal performance time. Desiccation was performed with Diquat (2 l ha-1) applied at 7-day intervals from the end of flowering to harvest maturity. There were 6 treatments and control (untreated). Germination was tested 2 and 21 months after harvest. The highest germination was recorded when the desiccation was performed 35 days after flowering. Results showed that optimal moment for applying chemical desiccation when there is no negative effect on the seed germination is specific for each sunflower genotype. Seeds with a high level of germination (>90%) can be stored for 19 months without significant loss in germination, namely, it was not negatively affected by chemical desiccation. Seed moisture and growing degree-days can be used as reliable indicators for optimal desiccation time. Cilj rada bio je da se ispita uticaj hemijske desikacije na klijavost i čuvanje kvaliteta semena tri inbred linije suncokreta, kao i da se ispitaju mogući indikatori za optimalno vreme izvođenja desikacije. Za desikaciju je korišćen Dikvat (2 l/ha) u intervalima od 7 dana od završetka cvetanja do žetvene zrelosti. Obavljeno je 6 tretmana i kontrola. Klijavost je ispitivana 2 i 21 mesec nakon žetve. Najveća klijavost je zabeležena pri desikaciji 35 dana posle cvetanja. Rezultati pokazuju da je optimalni momenat za izvođenje hemijske desikacije, u kom nema negativnog uticaja na klijavost, specifičan za svaki genotip suncokreta. Seme visokog kvaliteta (>90%) moguće je čuvati 19 meseci bez značajnog smanjenja klijavosti, takođe, kod takvog semena nije bilo ni negativnog uticaja hemijske desikacije. Vlaga semena i sume aktivnih temperatura se mogu koristiti kao pouzdani indikatori optimalnog vremena za izvođenje hemijske desikacije.

Climate changes are one of the crucial issues of modern agriculture. These changes imply the increase in average temperatures and the frequent occurrence of temperature extremes. Such conditions are stressful to maize concerning the expression of its traits. This paper presents the analysis of the maize yield concerning the yield components, morphological and chemical traits under various agroecological conditions. The objective of the study was to evaluate variability of grain yield (GY) and yield components (ear length, EL and number of kernel rows, NKR), morphological traits (plant height, PH and ear height, EH), as well as chemical traits (the whole plant dry matter, PDM and the ear dry matter, EDM), and the effect of the environment on the intensity of expression of these traits. Fifteen genotypes developed by crossing of six inbred lines were used as a material in the study carried out during the two-year period in one location. Obtained results indicate that traits were more pronounced in maize hybrids than in the parental components including the lower variability of the traits expression. NKR did not affect the yield unlike the EL, whose coefficient of determination was R2 = 0.600 in both production years. The environmental effect was high for all traits except NKR. PDM and EDM were closely related to PH and EH, but they also depended on the EL and the NKR. Variations in meteorological conditions have a significant impact on the final goal of the production, the yield. Prevailing agro-ecological conditions should be a major guideline in planning the crop production and selecting the genetic material.

The ongoing climate change, coupled with the transformations of cultivation practices and land use in sole crop production, may cause a significant decline in sunflower yield stability. Considering that the sunflower is the third oilseed in the world oilseed market, with 45 million tons of grain per year, and that it takes the fourth position in the vegetable oil market, the implementation of sustainable growing technology in sunflower production is a necessity. Considering the above, the goal of the research was to analyze and recommend sustainable sunflower production technology in intercropping systems. A four-year trial was conducted in Serbia’s agroecological conditions (45∘34′23.2′′ N 19∘86′18.9′′ E) using a split-plot design. Two oil-type hybrids and one confectionary-type hybrid were intercropped with common vetch, red clover, and alfalfa. Common vetch × sunflower intercropping resulted in the decrease in almost all sunflower traits. Moreover, sunflower × alfalfa intercropping proved to be the most appropriate. The yield of NS Gricko and Rimi PR were statistically on the same level with sole cropping, while alfalfa biomass had better results after alfalfa was intercropped with NS Gricko, as compared to sole cropping. In view of the prevailing belief that yields are more stable in intercropping than in sole cropping, further research is needed in this respect, in addition to further research of the time and method of sowing.

Seed treatments with zinc, boron, biostimulant Coveron and MIX (zinc + boron + Coveron) were applied to three lettuce and three celeriac cultivars. Seeds of three wheat cultivars were treated under laboratory conditions with Trichoderma harzianum and eight Bacillus spp. Seed germination, seedling growth, and the presence of the following pathogens were determined: Fusarium sp., Alternaria sp., Penicillium sp., and Mucor sp. The Coveron treatment was the most effective on lettuce seeds tested in the germination cabinet. Seed germination was higher by 4% than in the control. Alternatively, germination of seeds treated with boron in the greenhouse was higher by 12% than in the control. The Coveron treatment had the highest effect on the shoot length, which was greater by 0.7 and 2.1 cm in the germination cabinet and the greenhouse, respectively. This treatment was also the most effective on the root length. Zn, B, and MIX treatments increased celeriac seed germination by 14% in the germination cabinet. The Zn treatment was the most efficient on seeds tested in the greenhouse. The germination was higher by 15%. A significant cultivar × treatment interaction was determined in both observed species under both conditions. The maximum effect on wheat seed germination (8%) was achieved with the T. harzianum treatment in the Salazar cultivar. A significant interdependence (p ≤ 0.01 to p ≤ 0.001) was established between seed germination and the seedling growth. The interrelationship between seed germination and pathogens of all cultivars was negative.

The paper presents the three years efficiency results (2011-2013) of the work and ways of harvesting (three types of mowers, with or without spreading the forage) influencing the quality of the forage (the content of crude protein and crude fiber) during the three days drying process. These results indicate that the harvesting method can strongly affect the work efficiency, energy consumption, the forage drying intensity and the quality of forage used for hay and haylage preparation. The best quality of the forage was achieved when a drum rotating mower POTTINGER CAT 185 was used. Negative correlation was found between content of crude protein and the crude fibre content, depending on the type of mower, varying in the range between r= -0.978 and r= -0.882 (POTTINGER CAT 185 r= -0.882 P lt = 0.05 to JF STOLL SB 200 r= -0.978 P lt = 0.001).

The European corn borer (ECB) Ostrinia nubilalis and Fusarium ear rot Fusarium spp. pose a continuous threat to maize production worldwide. There are several reports indicating that ECB damage to maize ears promotes Fusarium ear rot infection. The aim of this study was to monitor the influence of different insecticide treatments (a.i. chlorantraniliprole, indoxacarb, and chlorantraniliprole+lambda-cyhalothrin) on the ECB and fungal diversity on maize kernels in the field in a four-year trial (2013-2016). A total of 16 different fungal genera were isolated from maize kernels, and Fusarium species were confirmed to be the dominant pathogens, present in all treatments, throughout the four years of experiments. The incidence of Aspergillus spp. and Penicillium spp. was established to be low. Apart from Fusarium species, the most frequent genera were: Aspergillus spp., Mortierella spp., Mucor spp., Penicillium spp., Acremonium spp. and Rhizopus spp. Treatments with chlorantraniliprole and chlorantraniliprole+lambda-cyhalothrin showed higher efficacy, though not statistically significant, compared to indoxacarb, in reducing the number of ECB larvae and damage they cause. However, no direct effect on the number of isolated fungal genera has been observed in any of the three insecticide treatments. Kukuruzni plamenac (Ostrinia nubilalis) i fuzariozna trulež klipa (Fusarium spp.) pričinjavaju najveće štete u proizvodnji kukuruza. Postoji nekoliko istraživanja koja su potvrdila da, svojom ishranom, larve kukuruznog plamenca doprinose i razvoju fuzarioznih plesnivosti na oštećenjima zrna i klipa kukuruza. Stoga je glavni cilj ovih četvorogodišnjih istraživanja bio da se odredi efikasnost insekticidnih tretmana na smanjenje brojnosti larvi kukuruznog plamenca, kao i njihov uticaj na diverzitet gljiva na zrnima kukuruza. Ukupno 16 različitih rodova gljiva je identifikovano u uzorcima zrna kukuruza tokom četvorogodišnjeg perioda istraživanja. Fusarium vrste su registrovane u svim tretmanima tokom sve četiri godine istraživanja, i svojim procentualnim učešćem potvrdile da su najdominantniji patogeni zrna kukuruza. Pojava vrsta rodova Aspergillus i Penicillium u uzorcima zrna kukuruza je bila slaba. Vrste koje su često bile registrovane zajedno sa Fusarium spp. na zrnima kukuruza su bile: Aspergillus spp., Mortierella spp., Mucor spp., Penicillium spp., Acremonium spp. i Rhizopus spp. Tretmani sa hlorantraniliprolom i hrorantraniliprol+lambda-cihalotrinom su pokazali veću efikasnost, ali ne i statistički značajnu, u odnosu na indoksakarb u smanjenju broja larvi kukuruznog plamenca. Takođe, i pored manjeg broja vrsta registrovanih u ova dva tretmana, nije uočen statistički značajan uticaj ni jednog ispitivanog tretmana na broj rodova gljiva prisutnih u uzorcima.

Sepsis is a life-threatening condition characterized by an acute cytokine storm followed by prolonged dysfunction of the immune system in the survivors. Post-septic lymphopenia and functional deficits of the remaining immune cells lead to increased susceptibility to secondary infections and other morbid conditions causing late death in the patients. This state of post-septic immunoparalysis may also influence disorders stemming from inappropriate or overactive immune responses, such as autoimmune and immunoinflammatory diseases, including multiple sclerosis. In addition, ongoing autoimmunity likely influences the susceptibility to and outcome of sepsis. This review article addresses the bidirectional relationship between sepsis and multiple sclerosis, with a focus on the immunologic mechanisms of the interaction and potential directions for future studies.

Maize seed is characterised by a variety of shapes and sizes, which directly or indirectly affect physiological properties. The aim of the present study was to establish the significance of the seed shape and size on the first count and germination. The following eight hybrids were used as a seed material in the trial: ZP196, ZP260, ZP341, ZP360, ZP434, ZP677, ZP684 and ZP704. The traits of seed shape and size were observed under laboratory conditions. The seed mass, the first count and germination were determined. The genotype and the seed fraction significantly affected the variance of seed mass formation, R2≥0,911, while the significance of these factors for the first count and germination was small with the coefficient of determination of R2≤0.129 and≤0.298, respectively. The first count and germination were high (above 95%) in all sizes and all shapes of seeds. The lowest first count and germination were detected in SO (94.2; 95.1, respectively), while the highest values were determined in SSP (97.1; 98.0, respectively). The importance of seed germination was determined by the analysis of variance (p

The aim of this study was to establish the effect of different seed treatments on germination , and the growth of the embryonic stem and the radicle of tomato and pepper varieties. Four treatments were used in the study: MIX [(Coveron + zinc (Zn 0.5%) + boron (B 0.025%)]; Coveron; zinc (ZnSO 4 , Zn 0.5%) and boron (B 0.025%). The treatments were applied on seeds of following four pepper varieties: Šorokšari, Somborka, Kraljica, and Mirtima and three tomato varieties: Rio Grande, Saint Pierre, and Tomato apple of Novi Sad (Novosadski jabučar). Germination and the growth increase of both the embryonic stem (cm) and the radicle (cm) were observed in the germination cabinets and pots containing soil in two laboratories – locations. Af- ter the treatment applied to the pepper seeds and testing in the laboratory germination cabinet the following was established: i) the maximum increase in germination of 90% was when the MIX and Zn treatment was applied to seeds, ii) the growth increase of embryonic stems of 2.7 cm was recorded when the MIX treatment was applied, iii) the growth increase of radicles of 1.7 cm was gained when the Coveron and MIX treatment was applied. Tests performed in pots showed that Coveron was the most efficient treatment. Treatments on tomato seeds during the seed testing in the germination cabinet provided: vi) the germination increase of 13% with the MIX treatment, vii) the growth increase of the embryonic stem of 2.6 cm with the same treatment, vii) the growth increase of 1.7 cm of radicles. Coveron was the most efficient treatment in tests in pots.

Plasmids are autonomous episomally replicating genetic elements, which carry backbone genes important for the replication and maintenance within their host, and accessory genes that might confer an advantage to their host under specific selective pressure in its ecological niche. The genome of dairy isolate L. lactis subsp. lactis bv. diacetylactis S50 was sequenced using the PacBio SMRT Cell Seq-RSII platform and revealed to possess one of the largest plasmidomes among L. lactis strains studied so far, harboring six plasmids: pS6 (5553 bp), pS7a (7308 bp), pS7b (7266 bp), pS19 (19,027 bp), pS74 (74,256 bp) and pS127 (127,002 bp) in total representing 8.9% of genome size (240,412 bp). Based on predicted plasmid replication proteins and origins it appears that all six plasmids replicate via the theta-type mechanism. The two the largest plasmids (pS74 and pS127), carry a number of genes known to be important for growth and survival in the dairy environment. These genes encode technological functions such as bacteriocin production, protein degradation, magnesium and cobalt/nickel transporters, selenium binding, exopolysaccharides (EPS) production, bacteriophage and stress resistance. Beside genes for replication, the small plasmids (pS6, pS7a, pS7a, and pS19) also carry genes important for mobilization and host survival such as type I restriction-modification (R-M) system, metal transporters, enzymes and transcriptional regulators. All plasmids in S50 strain are mobilizable, containing an oriT sequences, while pS127 is self-conjugative and allows for mobilization of the other plasmids. Small plasmids are prone to structural and segregational instability, while pS127 appeared to be segregationally stable thanks to the possession of two partition systems. The main characteristic of plasmid pS74 is EPS production, while plasmid pS127 is characterized by proteinase and multiple bacteriocins, tra locus, phage abortive systems and metal transporters. In addition to LcnA and LcnB, plasmid pS127 encodes several bacteriocin-pheromone molecules and a new bacteriocin named LcnS50, with narrow spectrum of action limited to lactococci, that has been successfully cloned and heterologously expressed.

Simple Summary Heterodera filipjevi, the cereal cyst nematode, is one of the most globally recognized and economically important nematodes on wheat. As some other cyst nematodes that are plant root parasites, the cysts of H. filipjevi survive in soil for years and shelter a large number of microbes. The aims of this study were to investigate the diversity of mycobiota associated with the cereal cyst nematode H. filipjevi, to infer phylogenetic relationships of the found mycobiota, and to explore the ecological connection between fungi and the field history, including the potential of fungi in bioremediation and the production of novel bioactive compounds. The study showed that the fungal species associated with the H. filipjevi cysts belong to diverse phyla, including Ascomycota, Basidiomycota, and Mucoromycota. The members of Ascomycota (Fusarium avenaceum, Sarocladium kiliense, Setophoma terrestris) are plant parasites, indicating that crops were host plants for fungal infection of recent origin. The members of Basidiomycota (Bjerkandera adusta, Cerrena unicolor, Trametes hirsuta, etc.) are wood-decay fungi, the presence of which in agricultural soil indicates that forests were the preceding plants. Abstract Cereals, particularly wheat, are staple food of the people from the Balkans, dating back to the Neolithic age. In Serbia, cereals are predominantly grown in its northern part between 44° and 45.5° N of the Pannonian Plain. One of the most economically important nematodes on wheat is the cereal cyst nematode, Heterodera filipjevi. Cysts of H. filipjevi survive in soil for years and shelter a large number of microorganisms. The aims of this study were to investigate the diversity of mycobiota associated with the cereal cyst nematode H. filipjevi, to infer phylogenetic relationships of the found mycobiota, and to explore the ecological connection between fungi and the field history, including the potential of fungi in bioremediation and the production of novel bioactive compounds. Cysts were isolated from soil samples with a Spears apparatus and collected on a 150-µm sieve. The cysts were placed on potato dextrose agar, and maintained for two weeks at 27°C. Following fungal isolation and colony growing, the fungal DNA was extracted, the ITS region was amplified, and PCR products were sequenced. The study showed that the isolated fungal species belong to diverse phyla, including Ascomycota, Basidiomycota, and Mucoromycota. Ascomycota is represented by the families Clavicipitaceae, Sarocladiaceae, Nectriaceae, and Phaeosphaeriaceae. Basidiomycota is represented by the families Cerrenaceae, Polyporaceae, Phanerochaetaceae, and Meruliaceae, and the order Cantharellales. The family Mortierellaceae represents Mucoromycota. The members of Ascomycota and Basidiomycota both depict the field history. Ascomycota indicate the fungal infection is of recent origin, while Basidiomycota point toward the preceding host plants, enabling the plant field colonization history to be traced chronologically.

Experimental autoimmune encephalomyelitis (EAE) is classically induced with complete Freund's adjuvant (CFA). The immune response against CFA has a confounding influence on the translational capacity of EAE as a multiple sclerosis model. Here, we compare clinical, cellular and molecular properties between syngeneic spinal cord homogenate (SCH)- and SCH + CFA-immunized Dark Agouti rats. EAE signs were observed earlier and the cumulative clinical score was higher without CFA. Also, a higher number of immune cells infiltrates in the spinal cords was noticed at the peak of EAE without CFA. High spinal cord abundance of CD8+CD11bc+MHC class II+ cells was detected in SCH-immunized rats. Myelin basic protein -specific response can be elicited in the cells from the lymph nodes draining the site of SCH immunization. This CFA-free EAE is a reliable multiple sclerosis model.

In this study, for the first time, the comprehensive analysis of antiproliferative and antioxidant activities of ramson, followed by the analysis of its associated microbiota and health-promoting effects of lactic acid bacteria (LAB), was performed. Ramson (Allium ursinum) is recognized as a medicinal plant with a long history of use in traditional medicine due to its antimicrobial and antioxidant activity. In this study the influence of in vitro gastrointestinal digestion on the cytotoxic activity of A. ursinum extracts against human malignant cell lines was demonstrated. Seven sulfur compounds, the degradation products of thiosulfinates, including diallyl disulfide were shown to inhibit proliferation of malignant cells by inducing accumulation within G2/M phase as well as to induce apoptosis through activation of caspase-3 and mitochondrial signaling pathway. Further, the A. ursinum microbiota, particularly LAB with potential probiotic effects, was analyzed by culture-dependent method and culture-independent method [denaturing gradient gel electrophoresis (DGGE)]. The obtained results revealed that the most abundant genera were Streptococcus, Lactobacillus, and Bacillus. The Lactobacillus genus was mainly represented by L. fermentum. The pulsed-field gel electrophoresis (PFGE) analysis revealed the presence of two PFGE pulsotypes. The probiotic potential of the strain L. fermentum BGSR163 belonging to PFGE pulsotype 1 and the strain L. fermentum BGSR227 belonging to the PFGE pulsotype 2 was characterized. The results revealed that both strains are safe for human use, successfully survive the simulated gastrointestinal conditions, have potential to transiently colonize the gastrointestinal tract (GIT) and have a protective immunomodulatory effect, inducing the production of proinflammatory cytokine IL17 and regulatory cytokine IL10, while decreasing the production of proinflammatory cytokine IFN-γ. In conclusion, the results of this study suggest that consumption of A. ursinum might have health-promoting properties, including anticancer effects, while L. fermentum strains isolated from A. ursinum leaves could be used as probiotics for human consumption.

Pseudomonas aeruginosa is one of the most common opportunistic pathogens that use quorum sensing (QS) system to regulate virulence factors expression and biofilm development. Delftia sp. 11304 was selected among 663 Gram-negative clinical isolates based on its QS inhibitory activity against P. aeruginosa MMA83 clinical isolate. Whole genome sequencing identified this isolate as D. tsuruhatensis and revealed genetic armamentarium of virulence factors and antibiotic resistance determinants. Ethyl acetate extract of D. tsuruhatensis 11304 culture supernatant (QSI extract) prevented biofilm formation of P. aeruginosa MMA83, but was unable to cause biofilm decomposition. QSI extract showed a synergistic effect in combination with meropenem and gentamycin, against P. aeruginosa MMA83. A dose-dependent reduction of the virulence factors: elastase, rhamnolipid and pyocyanin production by P. aeruginosa MMA83 and significant downregulation of lasI, lasR, rhlI, rhlR, pqs and mvfR expression were observed. Matrix-assisted Laser Desorption Ionization (MALDI) mass spectrometry of D. tsuruhatensis 11304 QSI extract revealed the presence of N-acyl homoserine lactones (AHL) with chain lengths of C12 to C18. The main ion peak was identified as N-octadecanoylhomoserine lactone (C18-HSL). Commercial C18-HSL (20 µM) reduced pyocyanin production as well as mRNA level of the lasI gene. A novel AHL species, dihydroxy-N-octadecanoylhomoserine lactone, was also described.

Burkholderia cepacia is well known as the causative agent of infections in humans where often shares niche with other pathogens, like Pseudomonas aeruginosa. Clinical isolate Burkholderia sp. BCC4135 was selected due to its strong quorum quenching (QQ) activity. Whole genome sequencing unveiled this isolate as B. cepacia with unique sequence type ST1485 and a myriad of genes belonging to resistome and virulome. Two QQ lactonases YtnP and Y2-aiiA originated from B. cepacia BCC4135 were cloned, expressed, and functionally characterized. They were active against a broad substrate spectrum of the N-acyl-homoserine lactones (AHLs). The YtnP lactonase was inactive, while Y2-aiiA was active against N-tetradecanoyl-dl-homoserine lactone (C14-HSL) which could imply the difference in their biological roles from the aspect of its quorum sensing (QS) autoregulation and interference with the QS systems of bacteria residing within the same niche. Both YtnP and Y2-aiiA were able to attenuate virulence potential of P. aeruginosa MMA83 clinical isolate declining its biofilm formation and virulence factors production. B. cepacia BCC4135 lactonases interfered with the las, rhl, and even pqs QS circuit of P. aeruginosa MMA83 transcription and the effect of combined enzymes was even more prominent. B. cepacia BCC4135 also employs the CepI/R QS system for governing its own virulence traits and possibly self-regulates the QQ/QS network through the different expression and activity of YtnP and/or Y2-aiiA. Our findings pointed out that BCC4135 lactonases could be exploited as an effective antivirulence drugs against P. aeruginosa and gave us a new insight into B. cepacia QQ/QS machinery.

A total of six tomato seed populations, collected in the autumn of 2016 at different locations in Serbia (West (3), South (1), North (1) and East (1)), were enrolled in this study. Seeds of the tomato populations considered were produced in the organic growing system in the area of Zaovice (76 m a.s.l., chernozem), Stara Pazova, Northern Serbia (44˚ 47 '19.6" N, 20˚ 27' 56.2" E) in 2017. Analyses of the germination parameters of the tomato seed populations examined (namely the germination energy, total germination, abnormal seedlings and dead seeds of the tomato seeds considered) indicated significant (r lt 0.01) differences according to the production year (A) and the tomato seed population (B). The impact interactions (r lt 0.01) of these factors (A × B) proved significant only relative to dead seeds. A highly significant positive correlation was found between the germination energy and the total germination of the seeds considered (r = 0.8711, p lt 0.001), as well as between the amounts of their abnormal seedlings and dead seeds (r = 0.92297, p lt 0.001). The germination energy and the total germination of the tomato seeds considered were found to decrease with the increasing seed age, in contrast to the numbers of their abnormal seedlings and dead seeds which continued to increase with seed aging. Seme šest populacija/sorti paradajza prikupljeno je u jesen 2016. sa različitih lokaliteta iz Srbije: zapadne (3 populacije), južne (1), severne (1) i istočne (1). Proizvodnja semena populacija paradajza u organskom sistemu gajenja izvedena je 2017. godine na lokaciji zapadne Srbije u Staroj Pazovi, potes Zovice (76 m n.v.) 44˚ 47 '19.6" N, 20˚ 27' 56.2" E. Cilj istraživanja je bio da se izvrši ocena uticaja godine i populacije na pokazatelje kvaliteta semena paradajza tokom 2017, 2018. i 2019.. Seme je čuvano u skladištu upakovano u pvc-kesama na temperaturi ispod 15ºC, RH 50%. Analiza energije klijanja, ukupne klijavosti, neneormalnih klijanaca i mrtvog semena paradjza pokazala je visoko značajne razlike (r lt 0,01) pod uticajem faktora godina (A) i populacija semena (B). Veoma značajna interakcija (r lt 0,01) ispitivanih faktora u pogledu mrtvog semena paradajza dobijena je samo kod međusobnog uticaja faktora A × B. Zabeležena je visoko značajna korelacija (p≤0.001) između energije klijanja i ukupne klijavosti, i između broja neneormalnih klijanaca i mrtvog semena. Energija klijanja i ukupna klijavost sa povećanjem starosti semena opadaju, dok broj nenormalnih klijanaca i mrtvog semena raste. Rezultati ovog rada treba da daju doprinos u shvatanju značaja autohtonih populacija/sorti ne samo paradajza, već i autohtonih sorti semena ostalog povrća; odnosno da ukažu na značaj očuvanja genetičkih resursa za organsku poljoprivredu.

This study aimed to analyze an impact of Lactobacillus salivarius BGHO1 and Lactobacillus reuteri BGGO6-55 supplemented to Artemia franciscana cultivation medium on biochemical profile of hatched nauplii. Impacts of different BGHO1:BGGO6-55 ratios (75:25, 50:50 and 25:75) and their total concentrations (250 and 500 g/kg of Artemia cysts) at different timings of bacteria application (pre- and post-hatching) were examined. The effects were evaluated by quantification of naupliar peptide, soluble protein, phospho-, neutral and unsaturated lipid content. Uni and multi-factorial analysis of variance (ANOVA) were used to estimate the effects of treatments relative to control and to model factor interactions, respectively Statistical analysis indicated that post-hatching application of 75:25 strain ratio at high dose was associated with an increase of neutral lipid amount. Furthermore, factor interaction profiling identified positive correlation of lactobacilli concentration with the level of free amino acids/short peptides, phospho-, neutral and unsaturated lipids, but only at 50:50 strain ratio combination. Application of lower lactobacilli dose at 75:25 strain ratio caused an increase of soluble protein and phospholipid amount. Hypothetically, graded response of Artemia nauplii to lactobacilli supplementation was induced with different BGHO1 doses. It assumedly ranged from stress-response protein synthesis at lower doses to membrane permeability alterations and triglyceride-mediated defense mechanism activation at higher BGHO1 doses. In contrast to lactobacilli supplementation after Artemia hatching, pre-hatching application of lactobacilli was not associated with an increase of Artemia nutritive profile, though again there was a positive association of lactobacilli concentration with nutrient amount. This implicates interference of lactobacilli with the hatching process. Results presented here facilitate the design of future studies aiming to modify Artemia nutritive profile in accordance with nutritional demands of cultivated fish species.

This paper presents the results of the cleaning tests of three different natural alfalfa seed lots and three different red clover seed lots performed using seed processing machines. The alfalfa (Medicago sativa L.) and the red clover (Trifolium pratense L.) are two of the most important perennial forage legumes in the Republic of Serbia. Alfalfa and red clover seeds for sowing should be clean, i.e. free from weeds and foreign matter, and of high germination and genetic values. Such requirements can only be achieved by cleaning, i.e. removing all undesirable impurities and poor-quality seeds. The following parameters important for the seed cleaning process and equipment were observed: seed purity, contents of weed and other crop seeds, inert substance content, amounts of processed seeds (kg), seed loss and processing output. The results obtained facilitate (depending on the amount and type of weeds and other impurities found in the alfalfa and red clover seed lots considered) the proper adjustment of the seed processing equipment to obtain the largest possible quantity of quality seeds that are in compliance with the laws and regulations of Serbia.

The ability of commercial bumblebees to transfer Clonostachys rosea spores as a biocontrol agent (BVT-CR7 strain) from the hive to sunflower heads for the control of Sclerotinia head rot was studied during 2016 and 2017 in field trials. The bumblebee hives were placed in isolation cages before flowering and allowed to adapt to the new environment. The bumblebee activity in 2016 was significantly lower in the confined space compared with colonies in the open field, while in 2017 their activity was similar at all sites. Artificial inoculation with Sclerotinia sclerotiorum ascospores was done at full bloom, and end of flowering. Floret samples collected at the end of the flowering period confirmed the presence of C. rosea in treated plants (7.4%). The presence of S. sclerotiorum was lower (up to 30%) than in the control heads, confirming efficacy of the treatment. Statistical tests showed that the difference in disease incidence was highly significant between successive evaluations. Mean disease incidence was 2, 13 and 57% when the biocontrol agent was present compared to 6, 26 and 68% in the controls. Seed samples showed that seed yield, 1000 seed weight, hectoliter mass and seed germination were increased in the treated plants for 11, 1, 3 and 6%, respectively. These results indicate that bumblebees can be used as efficient vectors of C. rosea spores to sunflowers for the suppression of Sclerotinia head rot and the added benefits of insect-mediated pollination.

Managing N, P and K inputs in semi-natural meadow production systems is important for achieving maximum yields in livestock farming. The objective of the present study was to estimate the effect of different NPK levels (N0P0K0, N50P50K50, N100P50K50, N100P100K100, N150P100K100 and N200P150K150 kg ha-1 yr-1 ) on the yield, quality and nitrogen nutrition index (NNI) in a grassland community of Agrostietum capillaris (semi-natural meadow) in western Serbia. The study was conducted during the seasons of 2005-2008. The values of the investigated parameters, except for the unit N uptake, were the highest in 2004/2005 due to favorable climate conditions. The levels of nitrogen significantly increased all of the studied parameters compared to the control treatment, except for unit N uptake. Mineral fertilizers at N200P150K150 provided the highest green forage yield (25.12 t ha-1 ), dry matter yield (8.12 t ha-1 ), crude protein yield (876.3 kg ha-1 ), nitrogen uptake (140.2 kg ha-1 ) and nitrogen nutrition index (70.2%), and the lowest unit N uptake (0.0022 kg N kg DMY-1 ). The use of mineral fertilizers increased green forage yield, dry matter yield and crude protein yield, increasing fertilizer from lowest to highest rate increased fresh and dry matter yield, as well as protein yield. Based on the results of the study, monitoring of nutrition indices would be necessary in order to increase productivity and economic benefits. Unošenje N, P i K inputa u poluprirodnim livadama je vaţno za postizanje maksimalnih prinosa na stoĉarskim farmama. Cilj ove studije bio je procena uticaja razliĉitih nivoa N, P i K (N0P0K0, N50P50K50, N100P50K50, N100P100K100, N150P100K100 and N200P150K150 kg ha-1 yr-1 ) na prinos, kvalitet i indeks ishrane azotom (NNI) u livadskoj zajednici Agrostietum capillaris (poluprirodna livada) u zapadnoj Srbiji. Studija je sprovedena tokom perioda 2005-2008. Vrednosti ispitivanih parametara, izuzev usvajanja azota po jedinici mase, bile su najviše u 2004/2005 zbog povoljnih klimatskih uslova. Nivoi azota su znaĉajno povećali sve ispitivane parametre u poreĊenju sa kontrolnim tretmanom, osim usvajanja azota po jedinici mase. Đubrenje sa N200P150K150 obezbedilo je najveći prinos zelene krme (25.12 t ha-1 ), prinos suve materije (8.12 t ha-1 ), prinos sirovih proteina (876.3 kg ha-1 ), usvajanje azota (140,2 kg ha-1 ) i indeks ishrane azotom(70,2%), kao i najmanji (0,0022 kg N kg PSM-1 ). Upotreba mineralnih Ċubriva povećala je prinos zelene krme, prinos suve materije i prinos sirovih proteina, povećavanjem Ċubriva od najmanje do najveće koliĉine je povećalo prinos sveţe i suve mase, kao i prinos proteina. Na osnovu rezultata studije, praćenje indeksa ishrane azotom bi bilo neophodna mera u cilju porasta produktivnosti i ekonomske efikasnosti.

The paper presents the seed production technologies of maize sown on three different sowing dates. Seeds of three maize inbred lines (L1, L2, L3) were used as a seed material in the location of Zemun Polje in 2018. The objectives of the present study was to determine the importance of different sowing dates as a method to overcome stressful conditions caused by unfavourable environmental factors, as well as to point out to a significance of the seed size in sowing. Effects of the following factors were observed in relation to emergence and the maize grain yield: sowing date (SD), seed fraction (LF, SF and PF) and genotype (L). The gained results indicate that the lowest percentage of emergence was determined in the variant L2/SD2/SF (34%), while the highest grain yield was determined in the variant L3/SD1/ PF 8.86 t/ha. The standard deviation of the yield is the largest for variants with the highest yield U radu je prikazana tehnologija semenske proizvodnje kukuruza sa tri različita datuma setve. Kao semenski materijal koriščeno je seme tri smooplodne linije L1, L2, L3 na lokaciji Zemun Polje u 2018 godini. Cilj rada je bio utvrđivanje značaja primene različitih datuma setve kao način prevazilaženja stresnih uslova nastalih nepovoljnim ekološkim činiocima, kao i ukazati na značaj krupnoće semena u setvi. Efekti faktora :datum setve (SD), frakcija(LF, SF, PF) i genotip(L) posmatrani su u odnosu na broj k lijalih semena posle setve i prinos zrna kukuruza. Rezultati ukazuju da je najniži procenat niklih semena bio u primenjenoj varijanti L2/SD2/SF (34%) i najveći prinos za varijantu L3/SD1/PF 8.86 t/ha. Standardna devijacija prinosa najveća je za varijante sa najvećim prinosom dok je varijansa nicanja semena obrnoto proporcijonalna. Efekti faktora na nicanje u proizvodnim uslovima i prinos zrna kukuruza su različitog značaja. Rok setve kao tehnološka mera u proizvodnji semenskog kukuruza ali i u merkantiln oj proizvodnji zrna vrlo je značajna za procenat niklih semena, kao i njegova interakciji sa frakcijom i genotipom. Ukupan doprinos faktora u varijansi nicanja semena je preko 50% R=0,678. Na varijansu prinosa efekat je slabijeg intenziteta i iznosi R=0,490. Razlike dobijene primenom različitih varijanti u proizvodnji semenskog materija ukazuju na značaj delovanja faktora i njihovo dalje istraživanje.

Multiple sclerosis (MS) is a chronic autoimmune disease of the central nervous system. The aim of this work was to study the probiotic effect of γ-aminobutyric acid (GABA)-producer Lactobacillus brevis BGZLS10-17 on experimental autoimmune encephalomyelitis (EAE), an experimental animal MS model. Clinical EAE symptoms were monitored in Dark Agouti (DA) rats treated with L. brevis BGZLS10-17 strain, or supernatant obtained from 48 h culture of L. brevis BGZLS10-17 cultivated in growth medium with or without GABA precursor monosodium glutamate (MSG). The results revealed that oral administration of L. brevis BGZLS10-17 alleviates the symptoms of EAE in DA rats. Namely, treatment with BGZLS10-17 and the supernatant of the strain cultivated in medium with MSG delayed the onset, shortened the duration, and reduced the intensity of the disease in the period when the EAE symptoms in controls were most pronounced. The probiotic treated animals were completely recovered after forty days, unlike the control animals. The results indicate that supplementation with live strain or with supernatant containing GABA produced by L. brevis BGZLS10-17 could alleviate the EAE symptoms. However, the use of L. brevis BGZLS10-17 in functional food as probiotic for autoimmune diseases should be tested in clinical trials. [Projects of the Serbian Ministry of Education, Science and Technological Development, Grant no. 173019 and Grant no. 173035]

Antidepressant fluoxetine (Flx) is the first therapeutic choice for the treatment of major depression (MD), however neuroanatomical spots of its action remain unclear. Immunohistochemical detection of c-Fos protein expression has been used for mapping activated neuronal circuits upon various stressors and drugs. We investigated the effect of 3 weeks of Flx treatment (15 mg/kg/day) on changes in neuronal activity, by mapping the number of c-Fos+ cells, in several brain subregions in adult male rats of control and following 3 weeks of chronic social isolation (CSIS), an animal model of depression. The aim was to identify brain subregions activated by vehicle or Flx treatment in both controls or simultaneously applied with CSIS. Flx prevented depressive- and anxiety-like behaviors in CSIS rats. In controls, Flx increased the number of c-Fos+ cells in the anterior/posterior piriform cortex (aPirCx, pPirCx), retrosplenial cortex dysgranular (RSD) and granular, c region (RSGc), dorsal hippocampal subregions (CA1d, CA2, CA3d, DGd), lateral habenula (LHB), paraventricular thalamic nucleus, posterior part (PVP) and lateral/basolateral complex of amygdala (LA/BL). CSIS-induced neuronal activation was observed in brain subregions implicated in mood and other mental disorders such as aPirCx, pPirCx, caudate putamen (CPu), acumbens nucleus shell (AcbSh), RSD, RSGc, DGd, PVP and LA/BL. Flx increased neuronal activation in both controls and CSIS rats in the CA1d, CA2, CA3d, PVP, LA/BL, while in striatum increased neuronal activation was observed only in CSIS. Our data identify activated CSIS-related brain subregions and/or Flx treatment, in which Flx increased c-Fos protein expression in CSIS rats.

The phenomenon of genotype by environment interaction (GEI) represents permanent interest for breeders, geneticists and biometricians with its practical and theoretical aspects. We investigated GEI for grain yield of medium early maturity maize (Zea mays L.) hybrids from the official variety trial network by the Department of Protection and Recognition of Varieties of the Republic of Serbia that includes experimental maize hybrids on eight sites over two years. Environmental variables explained 77.6% and 60.7% of the GEI variation for two consecutive years, respectively. Factorial regression combined with stepwise procedure revealed the model which includes variables precipitation in July (pr7), minimum temperature in May (mnt5), maximum temperature in May (mxt5) and insolation hours in April (sh4), in 2004, and environmental index (EI) and average temperature in September (mt9) in 2005, to be the most explanatory models in the region of Vojvodina (Serbia) in two consecutive years. These results provide a base for further research in GEI and stability analysis, and are a useful tool in characterizing the sub-regions of maize growing area and extending the existing results to new sites. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. 451-03-68/2020-14/200032]

Decoctions obtained from dried apple and pear fruits were subjected to in vitro digestion in the presence of a food matrix in order to determine changes in the polyphenol content and antiproliferative activity. The total phenolic and total proanthocyanidins contents were determined using spectrophotometrical methods, and the phenolic compounds were analyzed by RP-HPLC/DAD before and after digestion. Chlorogenic acid and phlorizin dihydrate were the major identified compounds. The addition of a food matrix immediately decreased the contents of individual and total phenolics. After digestion, they were slightly elevated but still lower than in the initial samples. Antiproliferative activity was investigated on human epithelial carcinoma cell line (HeLa), human colon carcinoma (LS174) and human foetal lung fibroblast (MRC-5) cells. The exhibited growth inhibition was not correlated with the content of phenolics in the tested samples, indicating that it could not be explained solely by their content. With the exception of the apple decoction effect on HeLa cells, the digestion process resulted in significant increases in the antiproliferative activity.

Association analysis or linkage disequilibrium mapping is a method for identification of quantitative trait loci (QTLs) in a panel of divergent unrelated individuals based on historical recombinations during a crop’s domestication and selection. It should account for the population structure, which can be the result of adaptation to local conditions or selection, to reduce the possibility of declaring false-positive associations. The aim of this study was to determine potentially significant and consistent associations between markers and agronomic important maize (Zea mays L.) traits using association analysis in a diverse breeding material that can be ultimately implemented in maize selection. To this end, 96 maize inbred lines were evaluated in field trials at three locations in Serbia for eleven agronomic traits and analysed with microsatellite markers. Twenty five microsatellites were used to assess the population structure using Bayesian model-based clustering method and to test the significance of associations between the markers and the traits with general (GLM) and mixed linear (MLM) models. The cluster analysis divided maize inbred lines in four subpopulations, corresponding to the BSSS (Iowa Stiff Stalk Synthetic), LSC (Lancaster Sure Crop), Iodent heterotic groups and exotic and independent germplasm. The models identified associations between twenty five microsatellite markers and eleven agronomic traits, resulting in 133 and 71 associations across the environments for GLM and MLM, respectively. Some of the identified marker-trait associations were significant and consistent in several environments. The associations stable in several environments were identified between the markers bnlg1067 and two flowering traits; nc005 and bnlg434 and plant height, bnlg434 and ear height; bnlg1643 and umc1127 and leaf number, bnlg1360 and ear diameter; umc1019 and umc1506 and number of rows per ear; bnlg2305 and bnlg1451 and ear length, and between bnlg1175 and thousand-kernel weight. The results of this study indicate that these microsatellites could be used in marker-assisted selection of inbred lines, after validation of the marker-trait associations and testing combining abilities of the inbreds during hybrid development. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. TR-31073]

Paper description: Light-induced changes in plant traits can result in light-differential herbivory. Usually plants in the shade are more damaged. In dioecious species plant sex can influence the outcome of plant-herbivore interactions, with male-biased herbivory suggested to be the rule. The simultaneous effects of light and plant sex on herbivory, plant morphological, defensive and nutritional quality traits were not previously explored. In shade tolerant species, herbivore pressure was higher at sun exposed sites. Light affected intersexual differences in defensive and nutritional quality traits. However these did not result in sex-biased herbivory. Plant morphological traits influenced herbivory rather than chemical defense. Abstract: Interactions between plants and herbivorous insects can be shaped by light environment, resulting in differential herbivory in sun and shade. In dioecious species, the combination of plant sex and light-induced changes in defense traits and nutritive value can alter the patterns of foliar damage. We explored the combined effects of light environment and plant sex on natural herbivore damage and plant traits in the dioecious understory forb Mercurialis perennis on Mt. Kopaonik (Serbia). The role of plant traits in predicting the extent of damage was examined as well. Male and female plants from contrasting light environments (shade vs . sun) were analyzed with respect to leaf damage, as well as plant morphological and biochemical traits (size, specific leaf area, carbon-based defensive compounds and nutritional quality). We found prominent differences in herbivore damage between sun and shade conditions, but not between the sexes. Plants from the sun-exposed site had a significantly larger leaf area removed. The specific leaf area co-varied with herbivore damage in an inverse fashion, while leaf nutritional value had a moderate effect. Contrasting light conditions influenced the patterns of intersexual differences in the contents of condensed tannins and soluble proteins, with females exhibiting higher trait values. We found that factors defining risk of consumption were related to plant morphological traits and nutritive value rather than to chemical defenses. https://doi.org/10.2298/ABS171207007S Received: December 7, 2017; Revised: January 16, 2018; Accepted: February 11, 2018; Published online: March 1, 2018 How to cite this article: Selakovic SD, Stanisavljevic NS, Vujic VD, Rubinjoni LZ, Jovanovic ŽS, Radovic SR, Cvetkovic DD. Light and sex interplay: differential herbivore damage in sun and shade in dioecious Mercurialis perennis . Arch Biol Sci. 2018;70(3):…

This research aimed to test the effects of lactobacilli, applied to cultured pike-perch, either through hydrolyzed OTOHIME fish diet, or through Artemia nauplii, on fish growth, microbiota balance and skeletal development. On the 12th Day Post Hatching (DPH) fish were divided into following treatment groups: two groups received the combination of OTOHIME and nauplii enriched either with Lactobacillus paracasei BGHN14 + Lactobacillus rhamnosus BGTH or with Lactobacillus reuteri BGGO6-55 + Lactobacillus salivarius BGHO1, and one group received OTOHIME hydrolyzed by BGHN14 + BGT10 and non-enriched nauplii. Control group received non-enriched nauplii and non-hydrolyzed OTOHIME. The treatment lasted 14 days and fish were sacrificed on the 26th DPH for the assessment of digestive enzyme activity and microbiota composition. Individual total lengths and individual body weights were recorded at the end of the treatments, on the 26th DPH, and also on the 45th DPH, in parallel with the evaluation of skeletal deformities and fish survival. Our results indicated positive effect of Anemia enriched with BGGO6-55 + BGHO1 on fish growth, skeletal development and trypsin to chymotrypsin activity ratio (T/C), as an indicator of protein digestibility. Hydrolysis of OTOHIME was also associated with better skeletal development, higher T/C values and lower levels of Aeromonas and Mycobacterium spp., which are important fish pathogens. Though additional testing in larger cohort studies is needed, these observations are promising in terms of usage of probiotics for improved environmentally friendly production of pike-perch in Recirculating Aquaculture System (RAS).

Plant-herbivore interactions can be mediated by plant apparency, defensive and nutritional quality traits that change through plant ontogeny, resulting in age-specific herbivory. In dioecious species, opposing allocation patterns in defense may lead to sex-biased herbivory. Here, we examine how onto stage and plant sex determine levels of herbivore damage in understory herb Mercurialis perennis under field conditions. We analyzed variation in plant size (height, total leaf area), physical (specific leaf area) and chemical (total phenolic and condensed tannins contents) defense, and nutritional quality (total water, soluble protein and nonstructural carbohydrate contents) during the shift from reproductive to post-reproductive stage. Furthermore, we explored correlations between the analyzed traits and levels of foliar damage. Post-reproductive plants had lower levels of chemical defense, and larger leaf area removed, in spite of having lower nutritive quality. Opposing patterns of intersexual differences were detected in protein and phenolic contents during reproductive stage, while in post-reproductive stage total leaf area was sexually dimorphic. Female-biased herbivory was apparent only after reproduction. Plant size parameters combined with condensed tannins content determined levels of foliar damage during post-reproductive stage, while the only trait covarying with herbivory in reproductive stage was total nonstructural carbohydrate content. Our results support claims of optimal defense theory – sensitive stage of reproduction was better defended. We conclude that different combinations of plant traits mediated interactions with herbivores in mature stages. Differences in reproductive allocation between the sexes may not immediately translate into different levels of damage, stressing the need for considering different ontogenetic stages when exploring sex bias in herbivory.

This study evaluated whether inert feed enriched with Lb. paracasei subsp. paracasei BGHN14 may be used as a weaning diet for first feeding pike-perch larvae. Three experimental groups were weaned from the start of exogenous feeding: two groups were given inert feed enriched with BGHN14 either via 12 h incubation with live BGHN14 cells or via coating with homogenized BGHN14 cells and one group was supplemented non-enriched inert feed. In all three groups Artemia was co-fed with inert feed during weaning. Control group larvae were fed Artemia exclusively during the treatment period. Treatment lasted fourteen days, starting from the 6th day post-hatch (DPH). Larval sampling was performed on the 20th DPH for gene expression and enzyme activity analysis. Larvae were also sampled on the 32nd DPH for morphometric and body composition analysis. Our results showed that weaning of first feeding pike-perch larvae was associated with an increase of fish condition (0.72 ± 0.12–0.77 ± 0.11 versus 0.67 ± 0.11 in controls), but it suppressed skeleton development, according to Col1 mRNA expression (1 ± 0.51–1.06 ± 0.36 versus 2.07 ± 0.53 in controls) and reduced fat deposition (1.25 ± 0.23–1.49 ± 0.33 versus 1.84 ± 0.31% in controls). This presumably reflected lower availability of soluble proteins in microdiet as opposed to live food, along with high leaching rate of amino acids from solid feed particles, as reported in our previous studies. However, skeleton differentiation was not impaired in group weaned on BGHN14 homogenate coated feed (Col1 mRNA expression: 2.68 ± 0.72), which was enriched in skeleton building and taste stimulating amino acids. These larvae were also presented with substantially higher length (15.28 ± 2.55 versus 13.93 ± 2.31 mm in controls) and weight (26.56 ± 13.83 versus 21.03 ± 11.25 mg in controls), which correlated with lower trypsin activity (1.06 ± 0.13 versus 1.43 ± 0.26 mU/mg of proteins in controls) and an increase of PLA2 to trypsin activity ratio (453.12 ± 109.36 versus 264.84 ± 69.03 in controls). Present study suggests that weaning of first feeding pike-perch larvae using BGHN14 homogenate coated microdiet supports skeleton development and improves fish growth.

Simple Summary The western honey bee is one of the most economically and ecologically important species currently facing serious challenges in its whole area of distribution. The honey bee is a highly diverse species with about 30 subspecies that are adapted to regional climate factors, vegetation, pests and pathogens. The local populations of honey bees are rapidly changing and their diversity is constantly manipulated by beekeepers through the import of foreign queens, selection and migratory beekeeping. This manipulation may lead to such changes that honey bees lose their ability to thrive in the areas that were previously suitable for their wellbeing. To see how this human interference changed the genetic variability of native honey bee populations from Serbia, we sequenced part of the mitochondrial genome and compared them with published sequences. Our results suggest that human influence significantly changes the natural composition of honey bees in Serbia and that the presence of some previously reported subspecies could not be confirmed. Abstract Local populations of Apis mellifera are rapidly changing by modern beekeeping through the introduction of nonnative queens, selection and migratory beekeeping. To assess the genetic diversity of contemporary managed honey bees in Serbia, we sequenced mitochondrial tRNAleu-cox2 intergenic region of 241 worker bees from 46 apiaries at eight localities. Nine haplotypes were observed in our samples, with C2d being the most common and widespread. To evaluate genetic diversity patterns, we compared our data with 1696 sequences from the NCBI GenBank from neighbouring countries and Serbia. All 32 detected haplotypes belonged to the Southeast Europe lineage C, with two newly described haplotypes from our sample. The most frequent haplotype was C2d, followed by C2c and C1a. To distinguish A. m. carnica from A. m. macedonica, both previously reported in Serbia, PCR-RFLP analysis on the COI gene segment of mtDNA was used, and the result showed only the presence of A.m. carnica subspecies. An MDS plot constructed on pairwise FST values showed significant geographical stratification. Our samples are grouped together, but distant from the Serbian dataset from the GenBank. This, with the absence of A. m. macedonica subspecies from its historic range of distribution in southern Serbia, indicates that honey bee populations are changing rapidly due to the anthropogenic influence.

C57BL/6, BALB/c and NOD mice are among the most frequently used strains in autoimmunity research. NOD mice spontaneously develop type 1 diabetes (T1D) and they are prone to induction of experimental autoimmune encephalomyelitis (EAE). Both diseases can be routinely induced in C57BL/6 mice, but not in BALB/c mice. Also, C57BL/6 mice are generally considered T helper (Th)1-biased and BALB/c Th2-biased mice. Having in mind increasingly appreciated role of gut associated lymphoid tissue (GALT) cells in autoimmunity, especially in relation to gut Th17 and regulatory T (Treg) cells, our aim was to determine if there are differences in proportion of CD4 + T cell populations in mesenteric lymph nodes and Peyer's p atches of these mouse strains. Lower proportion of Treg was observed in NOD PP, Th2 cells dominated in BALB/c mice in mesenteric lymph nodes (MLN) and Peyer's patches (PP), while Th1 cells prevailed in C57BL/6 MLN. Intradermal immunization of mice with complete Freund's adjuvant resulted in significant difference in Th cell distribution in GALT of NOD mice. Differences were less pronounced in C57BL/6 mice, while GALT of BALB/c mice was almost unresponsive to the immunization. The observed strain- and tissue-dependent changes in Treg proportion after the immunization was probably a consequence of different CCR2 or CCR6-related migration patterns and/or in situ Treg proliferation. In conclusion, NOD, a highly autoimmunity-prone mouse strain, exhibits more profound GALT-related immune response upon immunization compared to the strains that are less prone to autoimmunity. Immunology Letters (2017), 190: 282-288

Cucumber (Cucumis sativus) fruit has been used in cuisine worldwide, while its leaves are rich in immunomodulatory compounds. Cucumber leaf extract (CLE) was characterized by the predominance of triterpenoids cucurbitacins and significant levels of phenolics. Effects of CLE on CD4 + T helper (Th) cells and macrophages, as the major encephalitogenic cells in the autoimmunity of the central nervous system were investigated in our study. CLE potently inhibited production of major pathogenic Th cytokines: interferon-gamma and interleukin-17, as well as of nitric oxide and reactive oxygen species in macrophages. Antigen-presenting activity of macrophages and dendritic cells was also affected by CLE. The effects of CLE were co-incident with modulation of NFκB and p38 MAPK signaling. Concentrations of CLE used in vitro did not show toxic effects on zebrafish embryos. Moreover, CLE inhibited generation of encephalitogenic cells in vivo. These results demonstrate that CLE deserve further investigation on its anti-encephalitogenic therapeutic properties. Journal of Functional Foods (2017), 37: 249-262

Fluoxetine (Flx) is the principal treatment for depression; however, the precise mechanisms of its actions remain elusive. Our aim was to identify protein expression changes within rat hippocampus regulated by chronic Flx treatment versus vehicle-controls using proteomics. Fluoxetine-hydrohloride (15 mg/kg) was administered daily to adult male Wistar rats for 3 weeks, and cytosolic and nonsynaptic mitochondrial hippocampal proteomes were analyzed. All differentially expressed proteins were functionally annotated according to biological process and molecular function using Uniprot and Blast2GO. Our comparative study revealed that in cytosolic and nonsynaptic mitochondrial fractions, 60 and 3 proteins respectively, were down-regulated, and 23 and 60 proteins, respectively, were up-regulated. Proteins differentially regulated in cytosolic and nonsynaptic mitochondrial fractions were primarily related to cellular and metabolic processes. Of the identified proteins, the expressions of calretinin and parvalbumine were confirmed. The predominant molecular functions of differentially expressed proteins in both cell hippocampal fractions were binding and catalytic activity. Most differentially expressed proteins in nonsynaptic mitochondria were catalytic enzymes involved in the pyruvate metabolism, citric acid cycle, oxidative phosphorylation, ATP synthesis, ATP transduction and glutamate metabolism. Results indicate that chronic Flx treatment may influence proteins involved in calcium signaling, cytoskeletal structure, chaperone system and stimulates energy metabolism via the upregulation of GAPDH expression in cytoplasm, as well as directing energy metabolism toward the citric acid cycle and oxidative phosphorylation in nonsynaptic mitochondria. This approach provides new insight into the chronic effects of Flx treatment on protein expression in a key brain region associated with stress response and memory. (C) 2017 Elsevier B.V. All rights reserved.

The objective of this study was to investigate the influence of seed coat colour on the seed quality of four red clover cultivars (germination, dormancy (hard seeds), dead seeds and initial seedling growth (vigour)) produced in Serbia over a period of two years. The seeds analyzed were first separated visually into bright, dark and mixed seed groups, followed by subsequent digital colour measurements. The results obtained showed that the seed coat colour of red clover could be considered a good indicator of seed quality and seedling growth ability. The results furthermore showed that bright-coloured seeds of red clover cultivars indicated increased vigour and seed quality compared to other colours. The impact of the production area and year on the seed quality parameters was inconclusive. A high variability in hard (CV = 22.22 %) and dead seeds (CV = 40.18 %) was recorded between different seed lots of red clover. A lower variability was found in the germination parameter of bright- and mixed-coloured seeds (CV = 4.53 % and CV = 8.53 %). The seed coat colour could be an important factor in determining the quality of red clover seeds, potentially increasing germination simply by removing dark-coloured seeds.

The aim of this study was to determine the effect of different seed treatments on germination parameters of three maize genotypes under optimal and suboptimal temperature conditions. Seed was treated with recommended doses of three commercial pesticide formulations: metalaxyl-m 10 g/L + fludioxonil 25 g/L, metalaxyl 20 g/kg + prothioconazole 100 g/kg and thiacloprid 400 g/L. Testing was conducted at 25°C and 15°C. Results of the study indicate that there are differences in response of maize genotypes to applied seed treatments, as well as to a specific treatment at optimal and suboptimal temperatures. Some treatments, depending on the mixing partner and temperature conditions, can affect final germination. In other cases, germination rate can be accelerated or prolonged, but with no effect on final germination. In order to provide fast and uniform emergence under different temperature conditions, further examination of the response of maize genotypes to specific seed treatments would be beneficial. Cilj istraživanja je bio da se utvrdi efekat različitih tretmana na parametre klijanja semena tri hibrida kukuruza na optimalnim i suboptimalnim temperaturama. Seme je tretirano preporučenim dozama komercijalnih pesticidnih formulacija: metalaksil-m 10 g/L+ fludioksonil 25 g/L, metalaksil 20 g/kg + protiokonazol 100 g/kg i tiakloprid 400 g/L. Ispitivanja su vršena u pesku, na 25°C i 15°C. Rezultati ispitivanja ukazuju da postoji razlika u reakciji genotipova kukuruza na primenjene tretmane, kao i na dati tretman u uslovima optimalnih i suboptimalnih temperatura. Neki tretmani, u zavisnosti od preparata koji se mešaju i temperaturnih uslova, mogu uticati na konačnu klijavost. S druge strane, brzina klijanja može biti promenjena, ali bez uticaja na konačnu klijavost. Kako bi se obezbedilo brzo i ujednačeno nicanje u različitim temperaturnim uslovima, dalja analiza reakcije različitih hibrida na date tretmane bi bila od koristi.

Research with the objective of finding efficient (non residual) sterilants for the purpose of greater automatization in establishing aseptic in vitro tissue culture was conducted on cherry rootstocks CAB 6P and Gisela 6. Two ways of sterilization were examined (NaOCl and ozone O3) through survival rate on three types of explants; buds with primordial leaves, buds without primordial leaves and nodal non lignified segment with axillary buds. Ozone resulted with the high rate of survival (from 57 to 93%) regardless of the type of explants and the rootstock variety. By introducing a complete bud without destruction significantly resulted with the higher rate of survival (from 90 to 97%). Results indicate the potential for ozone sterilization of the starting material not only as an ostensible but the perforated (sub-cuticular transpiration) agent., Istraživanja s ciljem pronalaženja učinkovitih sterilizanata, u svrhu veće automatizacije u uspostavi aseptične kulture tkiva in vitro, provedena su na podlogama trešnje CAB 6P i Gisela 6. Ispitivana su dva načina sterilizacije (NaOCl i ozon O3), prateći stopu preživljavanja na tri tipa eksplantata; pupovi s primordijalnim lišćem, pupovi bez primordijalnoga lišća i nodijalni nelignificirani segment s aksilarnim pupom. Ozon je rezultirao visokom stopom preživljavanja (od 57 do 93%), bez obzira na vrstu eksplantata i varijante podloge. Uvođenjem cjelovitoga pupa bez destrukcije značajno je rezultiralo višom stopom preživljavanja (od 90 do 97%). Rezultati ukazuju na potencijal sterilizacije ozonom inicijalnoga biljnoga materijala, ne samo kao površinskoga, nego i penetrirajućeg (subkutikularnog) agenta.

This paper presents the results of an analysis of maize seed production. A total of six hybrids were selected from two maturity groups (FAO 300 and FAO 600) produced in three locations (L1, L2, L3). The following commercially important seed traits and their variability were analyzed: 1000-seed mass and germination. The average 1000-seed mass of all the parameters examined amounted to 342.8 g, whereas the minimum and maximum masses recorded were 285.7 g and 370.5 g, respectively. In the FAO 300 hybrids, a 60:30 ratio of large to small seed fractions was recorded. Conversely, this ratio in the FAO 600 hybrids was not uniform. Seed germination of the hybrids examined was high (above 90 %). The highest effect on trait variability was recorded in the hybrid combination (η2 = 0.889 - seed mass, and η2 = 0.456 - germination). Trait variations across locations and hybrid combinations are important for obtaining high yields and realizing the maximum potential of seed material. U radu su prikazani rezultati analize proizvodnje semenskog kukuruza. Iz proizvodnje su odabrani sledeći hibridi: ZP 333, ZP 341, ZP 360, ZP 600, ZP 606 i ZP 666; dve grupe zrenja (FAO 300 i 600), proizvedenih na tri lokaliteta (L1, L2, L3). Analize su rađene na osobine semena koje su važne komercijalne karakteristike - apsolutna masa semena i klijavost, a koje zavise od uslova proizvodnje i vremenskih uslova godine, te je njihova varijabilnost manja ili veća u zavisnosti od ovih faktora. Varijabilnast i korelacija dve pomenute osobine su posmatrani na semenu proizvedenom u 2015. godini. Seme je podeljeno prema masi na krupnu (8,5-11 mm) i sitnu frakciju (6,5-8,5 mm). Prosečna masa 1000 semena za sve posmatrane parametre iznosila je 342,8 g, minimalana 285,7 g i maximalna 370,5 g. Hibridi grupe zrenja 300 karakterisali su se većim procentom krupne frakcije: odnos za hibride iz ove grupe bio je 60% krupne i 30% sitne frakcije. Grupa zrenja 600 nema ujednačen odnos frakcija između hibrida. Klijavost za sve posmatrane hibride bila je visoka - iznad 90%, uz mali broj uzoraka čija vrednost je bila ispod standardizovanog procenta. Varijabilnost u ispoljavanju osobina između lokaliteta bila je veoma značajna (p lt 0,05). Najveći efekat na varijabilnost osobina imala je hibridna kombinacija (η2=0,889 na masu i η2=0,456 na klijavost) a zatim interakcija hibridne kombinacije sa lokalitetom (η2=0,621 na masu i η2=0,623 na klijavost). Frakcija nije imala statističke značajnosti u ispoljavanju klijavosti semena (η2=0,020). Variranje osobina prema lokalitetu i hibridnoj kombinaciji značajno je za postizanje visokih prinosa i ispoljavanje maksimalnog potencijala semenskog materijala.

Chronic social stress and/or pharmacological treatments differentially modulate the expression of c-Fos, a marker of neuronal activity, in subregions of the rat brain. Here, we examined the effect of the atypical antipsychotic Clozapine (Clz) (20 mg/kg/day for 3 weeks) on the neuronal activation pattern of c-Fos protein expression in stress-relevant brain subregions of adult male Wistar rats exposed to chronic social isolation (CSIS: 3 weeks), an animal model of depression and schizophrenia, and controls. The protein expression of c-Fos was also used to map neuronal populations in brain subregions activated by CSIS alone. Subregions which showed significantly increased c-Fos protein expression following CSIS included the retrosplenial cortex (RSC), (subregions:RSC granular cortex, c region (RSGc) and dysgranular (RSD)), dentate gyrus, dorsal (DGd), paraventricular thalamic nucleus, posterior part (PVP), lateral (LA)/basolateral (BL) complex of amygdala, caudate putamen (CPu) and accumbens nucleus, shell (AcbSh). Increases in c-Fos protein expression in the RSGc, RSD, DGd, PVP, LA/BL complex of amygdala and striatum (CPu, Acb Core (AcbC) and AcbSh) following Clz treatment in controls were found. Clz applied simultaneously with CSIS modulated neuronal activity in CPu, AcbC and AcbSh subregions compared to CSIS alone, increasing c-Fos protein expression. Furthermore, Clz revealed synergistic effects with CSIS in the CA1d and PVP. These identified neural circuits reflect brain subregions activated following CSIS and/or Clz administration. These data further contribute to the understanding of the effectiveness of Clz in the modulation of brain subregion activation in response to CSIS.

The aim of this research was to improve nutritive value of fishmeal-based feed by lactobacilli in order to achieve satisfactory nutrient availability needed to support fish development. Feed was solid-state treated at a laboratory scale with the combination of Lactobacillus paracasei subsp. paracasei BGHN14 and Lactobacillus rhamnosus BGT10 in different experimental settings, which included the variation of strain ratio, total lactobacilli concentration, percentage of moisture and duration of incubation. Short peptides, soluble proteins, phospho-, neutral and unsaturated lipids were quantified. Differences among treated and control feeds were evaluated by Student t-test, while Gaussian process regression (GPR) modeling was employed to simulate the incubation process and define the optimal treatment combination in the context of overall feed nutritional profile. Treatment duration was shown to be the critical determinant of final outcome, either as single factor or via interaction with strain ratio. Optimal nutrient balance was achieved with 12 h incubation period, 260% moisture, 75:25 and 50:50 BGHN14:BGT10 ratios and 200 mg of lactobacilli per g of dry feed. This study should serve as the basis for large-scale tests which would simulate on-farm production of both fishmeal-based and unconventional, lower cost aquafeed with added value.

Gut microbiota dysbiosis has been considered the essential element in the pathogenesis of multiple sclerosis and its animal model, experimental autoimmune encephalomyelitis (EAE). Antibiotics were administered orally to Dark Agouti (DA) rats early in their life with the aim of perturbing gut microbiota and investigating the effects of such intervention on the course of EAE. As a result, the diversity of the gut microbiota was reduced under the influence of antibiotics. Mainly, Firmicutes and Actinobacteria were replaced by Proteobacteria and Bacteroidetes, while decreased proportions of Clostridia and Bacilli classes were accompanied by an increase in Gamma-Proteobacteria in antibiotic-treated animals. Interestingly, a notable decrease in the Helicobacteraceae, Spirochaetaceae and Turicibacteriaceae was scored in antibiotic-treated groups. Also, levels of short chain fatty acids were reduced in the faeces of antibiotic-treated rats. Consequently, aggravation of EAE, paralleled with stronger immune response in lymph nodes draining the site of immunization, and increased inflammation within the CNS, were observed in antibiotic-treated DA rats. Thus, the alteration of gut microbiota leads to an escalation of CNS-directed autoimmunity in DA rats. The results of this study indicate that antibiotic use in early life may have subsequent unfavourable effects on the regulation of the immune system.