Heavy Metals Content in Selected Medicinal Plants Commonly Used as

For the majority of the world population medicinal plants represent the primary source of the health care. Therefore determining the content of heavy metals, accumulated in medicinal plants, is of high importance. Thus, the purpose of this research was to evaluate the content of heavy metals and two metalloids in selected medicinal plants (Matricaria chamomilla L., Melissa officinalis L., Mentha piperita L. and Foeniculum vulgare Mill.), that are traditionally used in alternative medicine. The plant materials were collected in a highly industrialized town (Pančevo). Plant analyses were done according to ICP methodology. The obtained results show that the contents of the potentially toxic elements in the investigated medicinal plant species were below the recommended limits and those medicinal plants from the studied growing site are suitable for preparation of teas and medicinal extracts

Procena uticaja navodnjavanja višegodišnjih zasada na stabilnost strukturnih mikroagregata i rizik od stvaranja pokorice

The soil structure is very dynamic measure, especially in the topsoil horizon, given that changes can occur under the influence of climatic factors, cultivated plants, amelioration measures and tillage. In the area of the Danube region, in September 2015, it was conducted the study on the effect of the perennial plants irrigation on the structural microaggregates stability and the risk of the soil crusting on three selected locations in agricultural soil samples taken from the surface layer (0-30 cm). In the analysed soil samples in disturbed state it was determined the ratio between the total content of particles of less than 0.002 mm (clay) in soil samples prepared with sodium pyrophosphate and the content of such particles in soil suspension prepared with water. The degree of the stability of microaggregates was expressed according to Vageler, in relation to their estimated stability index. The risk of the soil crust formation was determined according to a Van der Watt & Claassen pattern. The obtained results showed that the soil samples have stable to very stable degree of the structural microaggregates stability and that an irrigation did not significantly affect the analysed parameter. In the analysed samples the risk of the soil crusting is high to borderline.Struktura zemljišta je veoma dinamična veličina, naročito u orničnom horizontu, koji je i ispitivan, obzirom da do promena može doći pod uticajem klimatskih činilaca, gajenih biljaka, primenjenih meliorativnih mera i obrade. Na području Podunavske oblasti, tokom septembra 2015. godine, sprovedena su ispitivanja uticaja navodnjavanja višegodišnjih zasada na stabilnost strukturnih mikroagregata i rizik od stvaranja pokorice na tri odabrane lokacije u uzorcima poljoprivrednog zemljišta iz površinskog sloja (0-30 cm). U ispitivanim uzorcima zemljišta u poremećenom stanju, određen je stepen stabilnosti mikroagregata, izražen prema Vageler-u, na osnovu određenog odnosa između ukupnog sadržaja čestica manjih od 0,002 mm (gline) u uzorku zemljišta pripremljenog sa natrijum pirofosfatom i sadržaja tih čestica u suspenziji zemljišta pripremljenog sa vodom. Određen je i rizik od stvaranja pokorice, po obrascu Van der Watt & Claassena. Dobijeni rezultati pokazuju da ispitivani uzorci zemljišta imaju stabilan do vrlo stabilan stepen stabilnosti strukturnih mikroagregata i da navodnjavanje nije u znatnoj meri uticalo na ispitivani parametar. U ispitivanim uzorcima rizik od stvaranja pokorice je visok do graničan

Rapd fingerprinting of indigenous lysinibacillus fusiformis isolates from stabilized sludge and oil-polluted soil

The Lysinibacillus group are motile Gram-positive bacilli, with oval or spherical spores, oxidase and catalase positive, not grouped in strict chains, with strictly oxidative metabolism and very similar to Bacillus sphericus group. Two indigenous isolates of Lysinibacillus fusiformis from oil-polluted soil near gas station and stabilized sludge from petrochemical plant in Serbia (FOV – HIP “Petrohemija”, Pancevo) were tested on IAR and HMT and showed high sensitivity to neomycin, cephalexin and bacitracin, and were resistant to trimethoprim. Both isolates were tolerant to 100μg/ml of Zn and Co, 10μg/ml of Mo, but they differed in tolerance to 20μg/ml of Cd and 10μg/ml of Hg. The isolate from stabilized sludge showed moderate emulsification ability of xylol (E24 65.8) and mineral oil (E24 75.6). The isolate from oil-polluted soil showed very strong emulsification ability of xylol (E24 87.2) and moderate of mineral oil (E24 72.7). RAPD fingerprinting showed clear differences between the two Lysinibacillus fusiformis isolates

Characterization of indigenous bacillus isolates from stabilized sludge in petrochemical industry

Bacillus species are rod-shaped, endospore-forming aerobic or facultatively anaerobic, Gram-positive bacteria; in some species cultures may turn Gram-negative with age. The many species of the genus exhibit a wide range of physiologic abilities that allow them to live in every natural environment. The spores are resistant to heat, cold, radiation, desiccation, and disinfectants. Bacillus species are used in many medical, pharmaceutical, agricultural, and industrial processes that take advantage of their wide range of physiologic characteristics and their ability to produce a host of enzymes, antibiotics, and other metabolites. Certain Bacillus species are important in the natural or artificial degradation of waste products. We isolated 15 indigenous Bacillus isolates from stabilized sludge in petrochemical plant in Serbia (FOV – HIP “Petrohemija”, Pancevo) and investigated their morphological and biochemical characteristics, emulsification activity and sensitivity to antibiotics and heavy metals. In addition, we estimated the genetic diversity of isolates by RAPD and rep-PCR. Three of 15 isolates showed very strong emulsification ability of xylol (E24 from 95 to 100). Six isolates showed strong emulsification of mineral oil (E24 from 78 to 100). All isolates were tolerant to 100μg/ml of Zn and Co, 10μg/ml of Hg and Mo, while eleven isolates showed tolerance to 10μg/ml of Cd and six isolates to 100μg/ml Hg. Only one isolate was sensitive to trimethoprim (5 μg). All isolates were sensitive to bacitracin (40U), cephalexin (30 μg), clindamycin (2 μg) and neomycin (120μg), while five isolates were resistant to novobiocin (5μg) and two to bacitracin (40U). Based on PCR analysis, we assessed genetic similarity of investigated Bacillus isolates

Comparative analysis of pseudomonas population in oil-contaminated soils in Serbia and plant - pathogenic pseudomonas

Pseudomonas species are remarkable for their capacity to colonize almost all terrestrial and aquatic ecological niches. This genus includes species of ecological, economic and health-related importance. Although they are globally active in aerobic decomposition and biodegradation, Pseudomonas includes species pathogenic for humans, domestic animals and cultivated plants. The aim of this study was to identify members of the Pseudomonas species from oil-polluted soil, investigate their diversity and compare it to phytopatogenic strains isolated from host-plants near marked site. Isolates were described phenotypically according to carbon assimilation, fluorescence on King B medium and susceptibility patterns to 5 different heavy metals. In addition, they were characterized genotypically using plasmid profile and fingerprints obtained with the (GTG)5 primer. We have observed high heterogeneity within the Pseudomonas strains collected from oil- contaminated soils. Phenotyping and (GTG)5 pattern showed that similarities between strains ranged from 55% to 94%, with some strains showing high level of similarity with plant-pathogens

Molecular characterization of Cronobacter sakazakii isolated from different herbal teas and mixtures in Serbia

Cronobacter sakazakii je uzročnik humanih infekcija koje mogu prouzrokovati ozbiljne i čak fatalne posledice kod odojčadi i imunokompromitovanih odraslih osoba i dece. Zbog visoke tolerancije na osmotski stres, C. sakazakii je često izolovan iz dehidratisanih proizvoda, kao što su formule za bebe u prahu i biljni čajevi. Cilj ovih istraživanja je detekcija, identifikacija i molekularna karakterizacija Cronobacter sakazakii izolata iz formula za bebe i različitih biljnih čajeva uzorkovanih u prodavnicama u Srbiji ili testiranih pri kontroli izvoza. U svih 360 analiziranih uzoraka mleka u prahu za bebe nije detektovana kontaminacija bkterijom C. sakazakii. Medjutim, u 192 od 520 testiranih uzoraka čajeva utvrdjeno je prisustvo C. Sakazakii (37.1%). Visoka zastupljenost je detektovana u čajevima za decu - 51.6% i u “bebi” čajevima - 44.1%, a zatim u čajevima za medicinsku upotrebu -38%. Najveća grupa čajeva koji sadrže 1 biljnu vrstu (221) sadržala je 72 C. sakazakii - pozitivnih uzoraka (32.6%) i uključivala je čajeve od Sennae folium, Althaeae radix, Menthae piperitae folium, Chamomilae flos i Urticae folium. Molekularna karakterizacija izolovanih C. sakazakii iz različitih biljnih čajeva rep-PCR, RAPD i 16S rRNA analizom sekvenci ukazala je na visok stepen sličnosti soju C. sakazakii NCTC 8155. S obzirom da je ovo jedan od najpatogenijih kliničkih sojeva, ovi rezultati ukazuju na potrebu za povećanom kontrolom zbog mogućeg rizika koju ove namirnice predstavljaju za imunokompromitovane i zdrave korisnike, a posebno za bebe i decu.Cronobacter sakazakii is an important cause of human infections that can be serious and even fatal among premature neonates and immunocompromised adults or infants. Because of its high tolerance to osmotic stress, C. sakazakii is frequently isolated from dried foods, such as powdered infant formula and herbal teas. The aim of investigation was detection, identification and molecular characterization of Cronobacter sakazakii isolates from infant formula and various herbal teas collected from Serbian market and tested for import control. C. sakazakii was not detected in any of the 360 analysed samples of powdered infant formula. However, 192 out of 520 samples of herbal teas tested were positive for C. sakazakii (37.1%). The high prevalence was observed in teas for children (51.6%) and in “baby” teas (44.1%), followed by medicinal teas (38%). The largest one-herb-teas group (221 samples) contained 72 C. sakazakii-positive samples (32.6%) and involved Sennae folium, Althaeae radix, Menthae piperitae folium, Chamomilae flos and Urticae folium teas. Molecular characterization of isolated C. sakazakii from different herbal teas by rep-PCR, RAPD and 16S rRNA sequences analysis showed the high similarity to C. sakazakii NCTC 8155. Knowing this strain as one of the most pathogenic clinical strains, our results raise concern about the safety risks these foods pose to immunocompromised and healthy consumers, especially for babies and children

In vitro i in vivo efekti Pseudomonas spp. i Bacillus sp. na Fusarium acuminatum, Botrytis cinerea i Aspergillus niger na krastavcu

Cucumber (Cucumis sativus L) is an important member of the Cucurbitaceae family. Production of healthy nursery is necessary for high-quality production of this crop in greenhouses and in fields. With the idea of minimizing the use of pesticides and mineral fertilizers to preserve soil quality, we investigated the effects of plant growth promoting bacteria (PGPB) on growth promotion and protection of cucumber plants from phytopathogenic fungi. The effects of Pseudomonas spp. strains with different antifungal activities and Bacillus sp. Q10 strain with PGP activity were tested on cucumber plants. Antagonistic activity of Pseudomonas spp. against the growth of several phytopathogenic fungi isolated from cucumber: F. acuminatum, B. cinerea and A. niger, was observed. The influences of overnight cultures, supernatants and heat-stable antifungal factors were tested on the phytopathogenic fungi in vitro. Pseudomonas sp. K35 and K24 strains were more effective than P. chlororaphis Q16 and Pseudomonas sp. K27, showing 70-80% of fungal growth inhibition regardless of culture or fraction applied. The good antagonists that belong to pseudomonads and Bacillus sp. Q10 strain were used as mixtures for estimation of plant growth and health promoting effects on cucumber plants. Growth dynamics differed depending on the applied strain of Pseudomonas sp. The M3 treatment (a mixture of Bacillus sp. Q10 and P. chlororaphis Q16) stimulated the initial phase of growth, while M4 (a mixture of Bacillus sp. Q10 and Pseudomonas sp. K24) resulted in the maximal height at the final measurement. Significant differences in leaf and plant weight (M4), and leaf weight (M5, containing K35 strain) were found after the treatments. No significant differences in chlorophyll and NBI level were observed in any of the tested combinations. The obtained results suggested that M3 was suitable for stimulation of the early phase of cucumber growth, while the mixtures M4 and M5 improved plant protection and stimulated the later phases of cucumber growth.Krastavac (Cucumis sativus L) je važan predstavnik familije Cucurbitaceae, a proizvodnja zdravog rasada je nephodna za visoku produktivnost u plastenicima i na otvorenom polju. Da bi se umanjila primena pesticida i mineralnih đubriva i pri tome sačuvalo zemljište, ispitan je uticaj bakterija stimulatora biljnog rasta (PGPB) na rast biljaka i zaštitu od patogena. Primenjeni su sojevi Pseudomonas spp. sa različitim antimikoznim delovanjem i Bacillus sp. soj Q10 sa PGP aktivnošću. Praćena je antagonistička aktivnost Pseudomonas spp. na fitopatogene gljive izolovane sa biljaka krastavca: F. acuminatum, B. cinerea i A. niger. Testiran je uticaj prekonoćne kulture bakterija, supernatanta i termostabilnih antifungalnih faktora na rast ovih fitopatogena. Pseudomonas sp. K35 i K24 sojevi, koji pokazuju 70-80% inhibicije rasta gljiva bez obzira na primenjenu kulturu ili frakciju, efektivniji su od P. chlororaphis Q16 i Pseudomonas sp. K27. Sojevi Pseudomonas spp., koji su ispoljili visok stepen antagonizma, kombinovani su sa sojem Bacillus sp. Q10 i procenjen je uticaj na rast i zdravstveno stanje biljaka krastavca. U zavisnosti od primenjenog soja Pseudomonas sp., razlikovala se dinamika rasta krastavca. Tretman M3 - kombinacija Bacillus sp. Q10 i P. chlororaphis Q16 uticala je na početnu fazu porasta biljaka, dok je treatman M4 - kombinacija Bacillus sp. Q10 i Pseudomonas sp. K24, imala najviše uticaja na visinu biljke na kraju merenja. Uočene su značajne razlike za masu lista i masu biljke (M4) i masu lista (M5 sa sojem K35), dok sadržaj hlorofila i nivo NBI nisu imali značajne razlike kod svih ispitivanih kombinacija. Dobijeni rezultati ukazuju da je M3 pogodan za rane faze razvoja biljke, a kombinacije M4 i M5 pogodne su za zaštitu biljaka i u kasnijim fazama porasta biljaka. Potpuna ekspresija PGP svojstava za M4 i M5 može se utvrditi tek posle testiranja na siromašnim supstratima, koji će biti upoređeni sa rezultaima dobijenim na supstratima sa dovoljno hranljivih materija

Detection of toxin genes and RAPD analysis of bacillus cereus isolates from different soil types

The aim of this study was to detect genes for enterotoxins (hbla, entFM and bceT) and for emetic toxin (cer), to determine antibiotic resistance, and to estimate intraspecies diversity in B. cereus isolates by RAPD analysis. B. cereus was identified in 12 out of 117 indigenous Bacillus spp. using the classical microbiological methods and PCR. All isolates were resistant to penicillin and ampicillin, two to tetracyclin and four to trimethoprim-sulphamethoxazole. Also, all isolates produced inducible penicillinases and β-lactamase. Toxin genes were detected with PCR. EntFM and cer genes were present in all isolates, hbla in all, but two, and bceT in none. RAPD analysis was performed with four different primers, two of them designed for this study. The intraspecies diversity revealed 10 different patterns at the 90% similarity level. Two separate clusters were formed regardless of a soil type or utilization. The detection of genes encoding toxins in all B. cereus isolates indicated these bacteria as potentially pathogenic and seriously for human health. Regardless of a soil type or utilization, the RAPD analysis showed high intraspecies heterogeneity in B. cereus isolates. To the best of our knowledge, this is the first study to analyse the presence of entero- and emetic toxin genes and genetic heterogeneity in B. cereus isolates from different soil types and different soil utilization in Serbia. [Projekat Ministarstva nauke Republike Srbije, br. TR37006

Detection of Stolbur phytoplasma on blackberry: A new natural host in Serbia

During the late summer of 2007, a severe phytoplasma-like disease was observed for the first time in blackberry plants (Rubus fruticosus), commercial cv. Čačanska beztrna. Redness and downward rolling of leaves were symptoms observed in three localities in Central Serbia. The presence of Stolbur phytoplasma, belonging to the taxonomic subgroup 16SrXII-A, in diseased samples was confirmed by the PCR - RFLP analysis of 16S rDNA genes and elongation factor Tu (tuf) gene. A sequence analysis of the tuf gene confirmed homology with phytoplasmas stolbur tuf-type II detected previously in Italian grapevines and red clovers in the Czech Republic. This is the first report of Stolbur phytoplasma 16SrXII-A group tuf-type II on blackberries in Serbia.Simptomi nalik fitoplazmi prvi put su uočeni tokom leta 2007 g. na kupini (Rubus fruticosus) sorte Čačanska beztrna. Crvenilo i uvrtanje listova kao najčešći simptomi primećeni su na 3 lokaliteta u centralnoj Srbiji. Prisustvo Stolbur fitoplazme, koja pripada 16SrXII-A taksonomskoj podgrupi, utvrđeno je PCR - RFLP analizom 16S rDNK gena i gena za elongacioni faktor Tu (tuf) u uzorcima sa izraženim simptomima. Potvrđena je homologija dobijene sekvence tuf gena sa fitoplazmama Stolbur tuf- II tipa detektovanim na vinovoj lozi u Italiji i crvenoj detelini u Češkoj Republici. Ovo je prvi nalaz Stolbur fitoplazme 16SrXII-A grupe tuf- II tipa na kupini u Srbiji