Anastasia GARBUZNEAC, Svetlana BOORTSEVA, Tamara SIRBU, Maxim BYRSA, Natalia CHISELITA, Oleg CHISELITA
Abstract. The researches have been carried out to study the effect of 4-aminobenzoic acid on the growth, lipid synthesis, and fractional composition of lipids in the biomass of Streptomyces massasporeus CNMN-Ac-06 strain. Under laboratory conditions, during cultivation of S. massasporeus CNMN-Ac-06 strain, various concentrations of 4-aminobenzoic acid were added to the SP-I complex medium. The maximum amount of biomass in comparison with the control sample was obtained by cultivation of the strain on SP-I complex medium supplemented with 4-aminobenzoic acid in concentration of 1.37 g/L – 24.99±1.1 g/L, and the amount of total lipids in it was 1.96±0.08 g/L, by 32.28% more in comparison with the control sample. An increase in the phospholipid fraction was obtained on the SP-I complex medium supplemented with 4-aminobenzoic acid in a concentration of 2.74 g/L – 24.39% (when in control sample – 20.70%). Triglyceride and sterol fractions increased on the SP-I complex medium supplemented with 4-aminobenzoic acid in concentration of 1.37 g/L – 18.46% and 20.54%, respectively (when in control sample – 10.55% and 6.80%, respectively).
Key words: Streptomyces; complex medium; 4-aminobenzoic acid; biomass; lipids; lipid fractions.
Khadidja Hadjer MANAA, Abdenour AIT SAID, Lilya BOUCELHA, Ouzna ABROUS-BELBACHIR, Réda DJEBBAR
Abstract. Soil contamination by heavy metals is considered as one of the major environmental issues in recent years as it poses a threat to agricultural production due to their adverse effects on plant growth, food quality and environmental health. Pollution by heavy metals involves metal ions which are not always essential to plants as they affect growth and development. The application of a chelating agent, EDTA, increases the absorption of heavy metals by increasing their phytoavailability. Our results showed that lead and nickel have a negative impact on peas (Pisum sativum) as they affect their germination rate, plant growth and protein content. EDTA itself also causes the same negative effects as heavy metals since it represents a toxic xenobiotic substance. The combination of this chelator with Ni or Pb amplified the action of these heavy metals on the various parameters studied. The Oxidative stress is manifested through the accumulation of reactive oxygen species and MDA which is an indication of cellular membranes alteration. The enzymatic activities of catalase, ascorbate peroxidase and superoxide dismutase were stimulated by EDTA, heavy metals or a combination of the two. In addition, the activity of the glutathione S-transferase was greatly increased in response to EDTA and both heavy metals.
Key words: EDTA; lead; nickel; oxidative stress; Pisum sativum; pollution.
Tetiana IVANOVA, Mykola PATYKA
Abstract. The aim of research was to determine the taxonomic position of dominant microorganisms in the fruit body of mushroom of the representatives of the genera Pseudomonas based on phylogenetic analysis of the nucleotide sequence of the 16S rRNA gene. Identification the bacteria they used the analysis of the nucleotide sequence of 16S rRNA gene. Bacterial DNA was extracted from the suspension of bacterial cells using GeneJet Genomic DNA Purification Kit (Thermo Scientific), amplification of the 16S rRNA gene was performed with primers 27F (5’-AGAGTTTGATCCTGGCTCAG-3’), 1492R (5’- GGTTACCTTGTTACGACTT-3’). The taxonomic position is determined by dominant microorganisms in the mushroom based on phylogenetic analysis of the nucleotide sequence of the 16S rRNA gene. The nucleotide sequence of a fragment of the gene 16S rRNA of the mentioned above strains was registered in the international database GenBank (NCBI) with number TI-2019 (3). Obtained amplicon in size of ~1500 BP. The purified PСК-product was sequenced in two directions using a set of reagents «BigDye Terminator v 3.1 Cycle Sequencing Kit». Analysis of the isolated strains of Pseudomonas fluorescens or the similarity of the nucleotide sequences of the 16S rRNA gene revealed 99% similarity with sequences of typical representatives of the species concerned. Promising strains of Pseudomonas fluorescens can be successfully introduced in the metagenome of aboriginal groups of the substations as biological agents of microbial preparations. They can provide metabolic functions of biological systems of the Agaricus bisporus, and be practically valuable agents of bioprotector action, induction of systemic resistance of plants against bacterial pathogens.
Key words: sequencing; 16S rRNA; Pseudomonas fluorescens; phylogenetic identification.
Dinar MOHAMED OUALI, Rabéa GACEB-TERRAK
Abstract. Vitex agnus-castus L. (Lamiaceae) is a bushy shrub native to Asian and Mediterranean regions and is often used in traditional and popular medicine for its many biological properties and aromatic virtues. There are many studies devoted to the phytochemistry of species growing in endemic conditions, but little or no information is available on species growing in a subtropical desert climate. In this perspective, a comparative study with works carried out almost everywhere in the world, aims to analyze phenolic extracts and essential oil of leaves harvested in arid regions of the south-west of the Algerian Sahara. The phenolic extracts are characterized by a high content of anthocyanidins (6.1±0.07 mg·g-1 DM) and by the presence of 40 components detected by HPLC-DAD, among which five phenolic acids, two flavones and two flavonols are identified. The essential oil is characterized by chemical diversity of volatile substances, 68 (98.97%) are selected by GC-MS of which 52 (89.72%) are identified. Hydrocarbon compounds (25.03%) are less abundant than their oxygenated (64.69%). These were distributed as follows: 25 monoterpenes (32.12%), 3 sesquiterpenes (31.84%), 1 diterpene (0.16%), 2 norisoprenoids (0.29%) and a non-terpene oxygenated compound (0.28%). A unique compound characterizes the oxygenated sesquiterpenes, caryophyllene oxide, which represents one-third (31.5%) of the total area of the foliar essential oil; this molecule is absent or very rare in all the other essential oils indicated in literature. Considering these results, V. agnus-castus acclimatized in the arid Saharan zone, presents a particular chemotype compared to those of endemic populations.
Keywords: Vitex agnus-castus L.; Saharan arid zone; chemotype profile; analysis; comparative study.
Ibtissam LAIB, Ali Boutlilis DJAHRA
Abstract. This study aimed to investigate the antioxidant and antibacterial capacity of crude extract and bioactive compoundsfrom the aerial parts of Helianthemum lippii. The antioxidant activities of the crude extract and fractions ( dichloromethane, ethyl acetate, n-butanol, and anthocyanins) were assayed by Diphenyl 2-pycril hydrazine (DPPH), reducing power (RP), Total Antioxidant Capacity (TAC) and linoleic acid/β - carotene bleaching assay. Their antibacterial activity against six bacterial strains was evaluated by Agar diffusion and a modified broth macro-dilution method was used to measure the minimum inhibitory concentration (MIC). Its phenolic content was also verified by quantitative estimations The results demonstrated this plant and its fractions have an excellent capacity antioxidant, as dichloromethane fraction exhibited the highest antioxidant activity in tested DPPH, RP assays, and TAC with IC50 and effective concentration (EC50) values of 2.404 µg/mL, 0.592 µg/mL, 1.58 mg Ascorbic acid/g of extract respectively. In addition, dichloromethane fraction showed promising antibacterial activity, with strong efficiency against six strains bacteria with the inhibition zone reaching even 26.67 mm, while the MIC was very similar in all fractions and much better than the crude extract so that it ranged in values from 0.1563 to 0.9375 mg/mL. Finally, it is worth noting this plant and its fractions is a promising source of medicinal uses that could be applied in the treatment of many diseases caused by oxidative stress and bacterial infection.
Key words: Helianthemuml lippii (L.) Dum. Cours.; dichloromethane; ethyl acetate; n-butanol; antioxidant activity; antibacterial activity.
Kuldeep YADAV, Rameshwar GROACH, Kavita RANI, Rohit DUTT, Narender SINGH
Abstract. The isolation of intact and high-molecular mass genomic DNA is an initial and essential step for many molecular biology applications like restriction enzyme fingerprinting, polymerase chain reaction (PCR) etc. The presence of essential oils, polysaccharides, polyphenols and other secondary metabolites in the medicinal plants can hamper and reduce the yield and purity of extracted DNA. In this protocol, we describe simple modifications to the conventional cetyl trimethylammonium bromide (CTAB) method for reliable isolation of high molecular weight genomic DNA from Gloriosa superba - a potent medicinal plant. The purity of genomic DNA was confirmed by excellent absorbance (A260/A280) ratio of ~1.8 calculated from the spectrophotometric readings and the DNA concentration ranged from 75 to 308 µg/g of leaves. The PCR amplification from the isolated DNA was optimized to produce clear amplification in all the germplasm tested as mentioned in our earlier research. Besides being rapid, effective and technically easy, the protocol can also be used for the isolation of genomic DNA from other crop plants too.
Key words: Gloriosa superba; extraction; genomic DNA; PCR.
Ugochi Nneka KEMKA, Toochukwu Ekwutosi OGBULIE, Kanayo OGUZIE, Christogonus Oudney AKALEZI, Emeka Emmanuel OGUZIE
Abstract. Lignocellulosic biomass materials are ample, sustainable and renewable assets with potentially far-reaching amounts of energy. They are an interesting option to fossil fuels which allows manufacture of biofuels and other organic compounds. Only specific and targeted pretreatment processes can ensure effective degradation of these lignocellulosic biomass material. Pretreatment procedures are necessary because they are the specific solution to the recalcitrance of biomass materials. It directly influences the biogas production potential. These procedures consist of physical, chemical, physico-chemical, biological or nanotechnological methods; emerging combined methods of lignocellulosic biomass pretreatment equally exist. Toxic inhibitors make up part of the drawbacks faced during pretreatment due to the fact that they immobilize cellulase and impede the growth of fermentative microbial consortia. There is therefore a dire need to either remove these inhibitors or reduce their presence. Enhanced production of biogas is very much dependent of the type of biomass involved, the pretreatment method used and the volume of toxic inhibitors generated. This review intends to help grasp the potential of pretreatment procedures and combinations of the same and their impact on the production efficiency of biogas and help to outline approaches to handling the emission of toxic inhibitors during the process solving the challenges that have to do with inefficient pretreatment results.
Key words: lignocellulosic biomass material; biogas; pretreatment; toxic inhibitors.
Sana BENOSMANE, Amel ALAYAT, Hayat AYAD, Rym MERABET, Houria BERREBAH
Abstract. Paramecium tetraurelia is the most vulnerable freshwater ciliated protest to the pesticide-contaminated aquatic environment and is commonly used as a valuable biological model in studies investigating the monitoring of environmental quality. The present study was undertaken to screen the physiological, biochemical, and antioxidant responses of Paramecium tetraurelia. exposed to chlorpyrifos-ethyl (an insecticide). Paramecia received increasing concentrations of chlorpyrifos ethyl (0.04, 0.05, 0.06 and 0.07 mg/L) for 24, 48, 72 and 96h. Results revealed concentration- and time-dependent variations in growth and response percentage in exposed cells. In addition, protein and malondialdehyde levels and catalase activity were significantly increased in a concentration-dependent manner in treated cells compared with controls. However, the enzymatic activity of acetylcholinesterase and glutathione level showed a concentration-dependent decrease in treated paramecium cells compared with controls. In conclusion, chlorpyrifos-ethyl, especially at higher concentrations, proved to induce marked changes in the physiological and antioxidant profiles of Paramecium tetraurelia. which are promised to be used as an efficient monitoring organism for environmental water pollutants.
Key words: insecticide; Chlorpyriphos-ethyl; Paramecium tetraurelia.; antioxidants; growth; Acetylcholinesterase (ACHE).
Soumaya BOUKERCHE, Nesrine GHERAIRIA, Azzedine CHEFROUR
Abstract. The seasonal variability of yield, chemical composition and antibacterial activity of Myrtus communis L. essential oils was investigated. The essential oils were extracted by hydro-distillation technique from leaves, berries, flowers and floral buds collected at three stages of development from northeast Algeria. The essential oil yield values ranged from 0.09 to 0.94 % (w/w). They depend significantly on the phenological stage and the plant part used for the extraction of essential oils. Myrtle leaf is the valuable organ for the essential oil production with a yield varied from 0.44 to 0.94 % (w/w); the highest yield was obtained at the flowering stage. The GC-MS analysis indicated that all essential oils of myrtle were dominated by monoterpenes, α-pinene (37.16-49.39 %) and 1,8-cineole (25.87-49.11 %), which were the main compounds found in essential oils at each harvest period. The antibacterial activity was evaluated using the disk-diffusion method against Escherichia coli, Extended- Spectrum- β-Lactamase, Carbapenem-Sensitive Klebsiella pneumoniae, Staphylococcus aureus, Enterococcus faecalis, and Salmonella spp. The essential oil of Myrtle berries and leaves extracted before flowering stage showed the best inhibitory effect against all tested bacterial strains, with inhibition zones diameter rangingfrom 11.24-26.52 mm and 12.84-29.84 mm, respectively. Myrtus communis L. essential oils displayed however a variable degree of antimicrobial activity with an absolute action against Klebsiella pneumonia.
Key words: Myrtus communis L.; vegetative cycle; phenological stage; essential oil composition; antibacterial activity.
Nnenna Nnannaya OTI, Toochuwu Ekwutosi OGBULIE, Augusta Anuli NWACHUKWU, Mercy Ngozi MADUBUIKE
Abstract. Oil spills destroy farmlands with detrimental impact on agricultural crops, thereby deteriorating the health of humans and other animals that consume the crops from the polluted regions. Assays for this study were conducted between two soil groups polluted with 100 mL crude oil (A - soil polluted before planting [PB] and B - soil polluted two weeks after planting [PA]) within a period of 28 days from their pollution time; at Federal University of Technology, Owerri, Nigeria. The impact of oil spill on three edible plants (Allium cepa, Telfairia occidentalis and Zea mays) were evaluated through plant growth measurement, total chlorophyll test done by spectrophotometry of leaf samples and histo-anatomical investigations. Results obtained at the end of the experiment showed a decrease in plant height, leaf area and leaf number for all the polluted plants compared with their controls. However, plants grown on group A soil experienced delayed emergence and retarded growth, but survived better than plants grown on group B soil. The photomicrographs revealed alterations and anomalies in leaf structures for both polluted groups as compared to their control samples. A notable decline in total chlorophyll contents occurred amongst the plants in group B when compared to plants in group A. Therefore, this study deduced an elevated cytotoxic effect of crude oil in all the polluted crop samples, except for mild anomalies observed in the leaves of T. occidentalis in group A (UPB); which also showed no physical sign of crude oil pollution based on the non-observance of leaf yellowing, deformity or defoliation.
Key words: oil spill; foliar; cytotoxicity; anomalies; chlorophyll; histology.
Cristian Felix BLIDAR, Ildikó EGYED, Ioana Mihaela TOMULESCU, Ioana Maria TRIPON, Irina Neta GOSTIN
Abstract. Ipomoea tricolor Cav. (morning glory) is one of the most popular outdoor ornamental plants, cultivated for its beauty, such as gardens and parks. Due to these qualities, it has determined the production and cultivation of an increasing number of varieties regarding the flower’s colour and more recently it captured the attention for its potential pharmaceutical use. The main aim of this article is to investigate in vitro germination and the first stages regarding growth and development of morning glory by using four different culture media: solid (agarized) medium, liquid culture medium, double-layer culture medium with agarized basal layer and a liquid medium in the upper part and using Blidar filter paper bridges (FPB). All cultured media are based on Murashige-Skoog formula of 1962. The control was represented by the classical solid culture medium. The analysis of the results revealed that the best results were obtained on control culture medium and on FPB supporting the need for maximum possible aerated substrates for further growth and development of the morning glory.
Key words: Ipomoea; in vitro; seeds germination; physical nature of the substrate; filter paper bridge.
Asma ZOUBIRI, Ettayib BENSACI, Menouar SAHEB, Billal GASMI, El Hadi MAILBI, Yassine NOUIDJEM
Abstract. As a colonial bird and the most abundant gull in the Mediterranean region, the yellow-legged gulls Larus michaehellis is considered as an ideal species to investigate effects of environmental factors change. The breeding ecology of the Yellow-legged Gull was followed for the first time in the continental semi-arid area in Algeria, during two consecutive years 2014 and 2015. The breeding site (Ain Zada dam) located in the Central Hauts Plateaux around 60 Km far from the Mediterranean Sea. Comparatively to previews observations, a rapid and massive growth of this marine species in the continental regions of the country was observed. In the aim to analyze factors affecting nest-site selection some environmental parameters influencing breeding parameters were surveyed such as nest density, vegetation density and height. Our results, showed the direct effect of water availability on the nest site selection and laying period in the semi-arid area. The mean nest density found in the study area (0.73 ± 0.15 nest/m2) was higher than those reported in other Mediterranean colonies. However, nest density associated with vegetation density, but negatively correlated with island size.
Key words: semi-arid; breeding ecology; yellow-legged gull; environmental factors; nest-site.
Olga POSTOLACHI, Inna RASTIMESINA, Valentina VORONA, Elena NICOLAU, Elena CULIGHIN, Oleg BOGDEVICI
Abstract. In the presence of pesticides in the soil, there is a tendency for the predominance of only a few functional groups of microorganisms, thereby affecting not only the general structure of the community, but also different biological processes in the soil. The research presented in this article aims to monitor groups of microorganisms during the bioremediation of soil long-term and complex contaminated with persistent organic pollutants (POPs). The experimental scheme for bioremediation of soil polluted with POPs involves the alternation of anoxic-oxic conditions and the addition of a fertilizer in different concentrations. The microbiological analysis of the polluted soil before bioremediation revealed the predominance of three functional groups of microorganisms: bacteria that assimilate mineral nitrogen, oligonitrophilic bacteria and ammonifying bacteria. Small amounts of micromycetes and Azotobacter and total lack of actinomycetes in the polluted soil were determined. In the experimental variants with added fertilizer, after five cycles of bioremediation, the microbial biomass increased significantly (by 15 and 155 times), and the degradation of pesticides was more intense.
Key words: soil pollution; bioremediation; microbial population.
Valerina SLANINA, Nina BOGDAN-GOLUBI, Ludmila BALAN, Elena-Simona NEAGU, Robert Marian RUGINESCU, Madalin Iancu ENACHE, Alexandr DORIF, Victoria SACARA
Abstract: Microbial diversity and enzymatic activity of bacterial strains isolated from water samples, bottom sediments and biofilms of lake were investigated and identified by PCR and sequencing of the 16S rRNA gene. Screening procedures included decimal dilutions performing with incubation at 37°C, isolation of pure non-pathogenic bacterial cultures, isolate cultivation on agar in Petri plates at 30°C and tests on enzyme production. Enzymatic activity of 65 selected strains (Bacillus, Planococcus, Micrococcus, Paenibacillus, Arthrobacter, Lysinibacillus, Bhargavaea, Peribacillus, Kocuria) was determined used tests on plates and by spectrophotometric approach. The isolates found to be a promising candidate for agriculture, textile, food and pharmaceutical application by showed enzyme-producing of amylase, lipase, cellulase, especially catalase.
Keywords: lake; bacteria; identification; enzymes; production; spectrophotometric assay
Wassila BOUGHANEM, Ibrahim BOUSSALHIH, Mohamed LAZALI
Abstract. Lentil (Lens culinaris Medik.) is a popular Mediterranean legume crop grown for its nutritious seeds and to improve soil fertility. Lentil yield is a critical and difficult trait to improve in crop genetics because it is influenced by various factors that negatively affect seed yields and quality traits. The main constraint to legume production and symbiotic nitrogen fixation is the soil’s insufficient phosphorus (P) availability. This P-deficiency also affects rhizobia multiplication in the rhizosphere, reducing the likelihood of infection and nodule growth. The primary goal of this study is to determine the degree of adaptation of lentils to Pdeficiency to select the best genotypes grown, considering growth rate, grain yield, and efficiency in the use of rhizobial symbiosis. In this context, we are investigating the behavior of some lentil genotypes grown in the Khemis Miliana (Algeria) area over two growing seasons, 2018 and 2019. The results show that yield values vary between genotypes, ranging from 500 to 1200 kg‧ha-1; specifically, the genotypes ’Large blonde and ’Ibla 1 recorded the highest yield over two growing seasons. Furthermore, efficiency in the use of the rhizobial symbiosis of lentils was higher for ’Djendel during the 2018 season (1.57 g shoot DW‧g-1 nodule DW) than for ’Large blonde (0.2 g shoot DW‧g-1 nodule DW). The genotypes chosen for their P usage efficiency had higher yields and were the best at dealing with low P availability in the field.
Key words: legume; phosphorus; N2 fixation; rhizobia; symbiosis.
Kajal CHAUHAN, Bindu BATTAN, Ashwani KUMAR, Sunita DALAL, Jitender SHARMA, Sulekha CHAHAL
Abstract. The present study was aimed at isolating halotolerant endophytic bacteria from three halophytic grasses, namely Sporobolus marginatus A. Rich., Urochondra setulosa (Trin.) C.E. Hubb and Leptochloa fusca (L) Kutnth. Out of total 56 bacterial isolates, 24 were screened from S. marginatus, 15 from U. setulosa, and 17 from L. fusca. The identified bacterial endophytes belong to diverse bacterial genera, i.e., Pseudomonas, Bacillus, Enterobacter, Microbacterium, Rhizobium, Chryseobacterium, Brevibacillus, Klebsiella, Proteus, Escherichia, and Agrobacterium. Among these, Pseudomonas, Bacillus, and Enterobacter were more prominent in all three halophytic grasses. S. marginatus seems to be more enriched as compared to U. setulosa and L. fusca in terms of the number of isolates. Root and node seem to be the preferred explants. The isolates were further tested for their salt tolerance potential, as a result of which 24 isolates belonging to Pseudomonas, Bacillus, and Enterobacter genera demonstrated continual growth up to 10% salt concentrations (NaCl), suggesting their significant salt tolerating capability. Thus, the three halophytic grasses seem to be excellent reservoirs of halo-tolerant bacterial endophytes. Pseudomonas, Bacillus, and Enterobacter were more frequent as well as more competent genera in dealing with salinity stress and could be exploited for their plant growthpromoting potential in bioremediation of extreme saline soils and to enhance crop yield under salinity stress.
Key words: salinity; halophytic grasses; endophytes; sustainability; halotolerant.
Monica Liliana MARIAN, Oana Elena MARE-ROȘCA, Bogdan VASILESCU, Daniel NĂSUI, Lucia MIHALESCU, Zorica Marcela VOȘGAN
Abstract: Biodiversity has become a priority in modern times. Over the past two centuries, the explosive development of human economy has led to the occupation of formerly natural areas for economic infrastructure or residential purposes. These replacements have resulted in the transformation of natural habitats into anthropized ones, disrupting the balance between species. Moreover, human activities have facilitated the migration of species from one continent to another, and the introduction of alien species to habitats different from their origin often has a significant impact on local biodiversity. This study aimed to investigate the incidence of alien species in three types of forest habitats (91E0*, 92A0 - riparian woods, and 9110 - beech forests) and three types of grassland habitats (6430 - Hydrophilous tall herb fringe communities of plain and of the montane to alpine, 6510 - lowland hay meadows, and R3803 - hillside hay meadows) within a set of protected natural areas in North-Western Romania (Central Europe). Out of the 23 species identified in habitats of community interest within the protected areas, 5 species are common to all of them. Among all the analysed habitat types, 91E0* riparian woods are the most affected due to human pressures, such as agricultural fragmentation, weakening of their structure, and the presence of road networks in their immediate vicinity. Beech forests, even though they have been mostly managed without considering their inclusion in Natura 2000 sites, show resistance to the spread of invasive alien species (IAS) due to their cohesive cenotic structure. For herbaceous vegetation, the pattern of IAS behaviour is similar - the most numerous and widespread IAS are found in narrow and ecotonal wet grasslands, while in well-connected, stable, and sustainably managed meadows, the incidence of IAS is minimal.
Key words: protected natural areas; habitats; invasive species; fragmentation; cenotic structure.