Zeitschriftenartikel zum Thema „Pasture plants Physiology“

AbstractGrasshopper density and species composition were determined from 1982 to 1985 in four 75-ha pastures in the northern mixed prairie region near Miles City, MT. Treatment in two pastures consisted of sagebrush removal and interseeding alfalfa and cicer milkvetch. Two pastures were not treated. The treated pastures were grazed only half as long as the untreated pastures but with twice the number of steers. Forage yield decreased in all pastures from 1983 to 1985 because of below-average precipitation and grasshopper density increases. In one pasture, 10% of the legume seedlings were destroyed by grasshoppers. The annual rates of increase in total grasshopper presence were similar in treated and untreated pastures. However, Melanoplus sanguinipes (F.), which eats forbs and is a strong flyer, increased more in treated than untreated pastures; six additional abundant species, all of which were grass feeders, did not increase more in treated pastures. Thus, treatments did not markedly affect total grasshopper population trends but did influence species composition and provided additional food plants for some species.

Pasture brome (Bromus valdivianus Phil.) has the potential to increase current levels of herbage production and pasture persistence in New Zealand dryland, well-drained soils. However, there is little literature on the effect of defoliation management on growth of this grass under contrasting soil-water restriction levels. The growth physiology and performance of pasture brome were evaluated in pots in a glasshouse. Defoliation frequency (DF) treatments were applied based on three different accumulated growing degree-days (AGDD): 250, 500 and 1000 AGDD (high, medium, and low DF). At end of the first growing cycle (1000 AGDD), water availability was restricted to 20–25% of field capacity (FC) in half of the pots, while the other pots were maintained between 80–85% FC. Total accumulated herbage mass was positively related with the low DF and well-watered conditions (p < 0.05). At the final harvest, plants subjected to low DF had greater root mass than high and medium DF (p < 0.05). At each harvest, the leaf regrowth stage (LS) for low DF was 3.5, while for high and medium DF, the LS was 1.5 and 2.0; respectively. Tiller water-soluble carbohydrates were highest at the low DF and under 20–25% FC. Regardless of soil-water conditions, defoliation at 3.5 LS supports production, enhancing survival during a drought.

The correction of soil acidity and the evaluation of the effects of correctives used are not frequent practices in pasture areas. The benefits of silicon on the physiology and biochemistry of pastures is a subject that has not been explored in great detail. The objective of this study was to evaluate the performance of Urochloa brizantha plants (cultivar Xaraés) submited to condictions of water deficity and silicate fertilization through physiological and biochemical parameters. The experiment was carried out in pots under greenhouse conditions..The experimental design was randomized blocks, in a 2 × 2 factorial arrangement, with eight replications. The treatments consisted of a combination of the presence and absence of silicon (by means of soil correction using agrosilicon and dolomitic limestone, to raise the base saturation to 50%) with 2 soil water conditions (40 and 80% of field capacity). The analyzes were performed to determine: net CO2 assimilation rate, stomatal conductance, transpiration rate, water use efficiency, superoxide dismutase and catalase enzyme activities, shoot dry matter ,neutral detergent fiber and acid detergent fiber determinations and leave protein. The application of silicon in Urochloa brizantha cultivar Xaraés has the potential of attenuate the water deficity, increasing the photosynthesis, plant dry matter and antioxidant enzymes activity.

Nitrogen fertilization is determinant for pasture productivity, as it results in increasing forage yield and is associated with the growth physiology of forage plants. An experiment was carried out in a greenhouse to evaluate the effects of nitrogen rates and times of application after cutting on Tifton 85 Bermuda grass (Cynodon spp.) tillering. Plants were grown in a Typic Quartzipsamment soil and nitrogen rates were 0, 80, 160 and 240 mg kg-1 of soil; times of application were immediately after and seven days after cutting. A 4 x 2 factorial experiment was set in a completely randomized block design (n= 4). Plants were evaluated in two sequential growth periods - 39 and 41 days. Nitrogen rates affected tiller density, considering the initial number of tillers in the second growth and the emerged and final number of tillers at the end of the growth periods. Tiller weight increased up to the nitrogen rate of 201 and 185 mg kg-1 of soil in the first and second growth periods, and the correlation coefficients between the final number of tillers and dry matter yields in the top part of the plant for these periods were 0.92 and 0.94, respectively. The rate of tiller development, evaluated in each of the seven-day periods, was affected by nitrogen rates and time of application. There was an effect of rates and time of application in the first and a significant interaction between rates and time in the second growth period.

Heat and drought are two major limiting factors for perennial pasture production in south eastern Australia. Although previous studies have focused on the effects of prolonged heat and drought stresses on pasture growth and physiology, the effects of short term recurring combined heat and drought stresses and the recovery from them have not been studied in detail. A controlled environment experiment was conducted to investigate the growth and physiological responses of perennial ryegrass (Lolium perenne L.), cocksfoot (Dactylis glomerata L.), tall fescue (Festuca arundinacea Schreb.) and chicory (Cichorium intybus L.) plants exposed to two consecutive seven day heat (control = 25/15 °C day/night; moderate = 30/20 °C day/night and severe = 35/30 °C day/night) and/or drought stresses each followed by a seven day recovery period. During the first moderate and severe heat and drought treatments, maximum photochemical efficiency of photosystem II (Fv/Fm), cell membrane permeability and relative leaf water content decreased in chicory and tall fescue compared to perennial ryegrass and cocksfoot. However, during the second moderate heat and drought treatment, all species showed less reduction in the same parameters suggesting that these species acclimated to consecutive moderate heat and drought stresses. Chicory was the only species that was not affected by the second severe heat and drought stress while physiological parameters of all grass species were reduced closer to minimum values. Irrigation mitigated the negative effects of heat stress by cooling the canopies 1–3 °C below air temperatures with the most cooling observed in chicory. All the species exposed to moderate heat and drought were fully recovered and those exposed to severe heat and drought recovered partially at the end of the experiment. These findings suggest that chicory may be a potential species for areas subject to frequent heat and drought stress.

Plant–animal interactions impact on all elements of pasture and animal performance in grazing systems. The quality of pastures for animals can be described in terms of feeding value (FV), which is a combination of feed nutritive value (NV) and voluntary intake. There are numerous complex interactions between plant physiology and pasture FV and NV. This review focuses on these interactions in four key areas (plant growth strategies, phenological development, pasture regrowth, and response to environmental stress), extracting key principles and illustrating how plant breeding or management may be used to manipulate such interactions to improve FV. The FV is low in pastures with native species that have evolved in nutrient-poor environments, especially if there are greater proportions of C4 than C3 species in the sward. Reproductive development of grasses and long grazing intervals (which affect stage of regrowth) reduce the proportion of leaf and increase stem or dead matter content in the sward. This is exacerbated by environmental stresses such as warmer temperatures and water deficit. Management decisions provide a means of manipulating many of these interactions to improve the FV of pasture, especially by improving soil nutrient status, using irrigation where possible, introducing exotic perennial pasture species such as perennial ryegrass, phalaris and tall fescue, linking the timing of grazing to stage of regrowth, and carefully managing post-grazing residual sward state. Likewise, plant breeding has focused on altering the flowering date of grasses, reducing aftermath heading, and reducing lignification within the plant to improve the FV of pasture for livestock.

When exposed to warmer, nonstressful average temperatures, some plant organs grow and develop at a faster rate without affecting their final dimensions. Other plant organs show specific changes in morphology or development in a response termed thermomorphogenesis. Selected coding and noncoding RNA, chromatin features, alternative splicing variants, and signaling proteins change their abundance, localization, and/or intrinsic activity to mediate thermomorphogenesis. Temperature, light, and circadian clock cues are integrated to impinge on the level or signaling of hormones such as auxin, brassinosteroids, and gibberellins. The light receptor phytochrome B (phyB) is a temperature sensor, and the phyB–PHYTOCHROME-INTERACTING FACTOR 4 (PIF4)–auxin module is only one thread in a complex network that governs temperature sensitivity. Thermomorphogenesis offers an avenue to search for climate-smart plants to sustain crop and pasture productivity in the context of global climate change.

Existem fortes evidencias da ocorrência de mudanças climáticas globais por causa do aumento de gases de efeito estufa, as quais provavelmente serão cada vez mais severas no futuro. Para enfrentar esse problema, as plantas, serão confrontadas com opções limitadas para evitar a perda de seu hábitat ou extinção: adaptar-se, migrar ou morrer. Em geral, o aumento da concentração atmosférica do CO2 atuando como “fertilizante” para a fotossíntese poderia aumentar o crescimento e a produção de biomassa das plantas. No entanto, o aumento da temperatura global poderia afetar negativamente a fisiologia e a produtividade vegetal, bem como provocar alterações nos padrões de precipitação com impactos graves sobre a produção agrícola e outros serviços ecossistêmicos. Para uma melhor estimativa dos impactos das mudanças climáticas na agricultura, pecuária, ecossistemas de florestas e pastagens serão necessários grandes avanços científicos. No âmbito da pesquisa, será necessária e prioritária a elucidação das interações entre o elevado CO2, temperatura, fertilidade do solo, disponibilidade hídrica e o efeito de poluentes como o ozônio sobre o crescimento, o rendimento e a produtividade das plantas. Efeitos das mudanças climáticas sobre o florescimento, viabilidade do pólen, polinizadores e produção de grãos também precisam ser esclarecidos. Qualquer avanço do melhoramento genético das plantas e uso da biodiversidade visando reduzir a sensibilidade à elevada temperatura ou melhorar a resposta ao CO2 será de grande utilidade. There is strong evidence that global climate change because of increase in the greenhouse gases are already occurring and will become increasingly severe. To address this problem, the plants will be faced with limited options to avoid losing their habitat or extinction: adapt, migrate or die. In general, the increase in atmospheric CO2 concentration acting as "fertilizer" for photosynthesis could increase growth and biomass production of the plants. However, the increase in global temperature could adversely affect the physiology and plant productivity and cause changes in rainfall patterns with serious impacts on agricultural production and other ecosystem services. A better estimate of the impacts of climate change on agricultural, forest and pasture ecosystems will require urgent scientific advances. It will be necessary to prioritize the elucidation of interactions between elevated CO2, temperature, soil fertility, water availability and the effect of pollutants such as ozone on growth, yield, and productivity of plants. Effects of climate change on flowering, pollen viability, pollinators and yield will need to be elucidated. Any advancement of plant breeding and use of biodiversity for reducing the sensitivity to high temperature and improve the response to CO2 will be very useful. Keywords: global climate change, warming, biodiversity, agriculture, plant productivity.

The P efficiency of Italian ryegrass (Lolium multiflorum Lamk. cv Grasslands Tama) and phalaris (Phalaris aquatica L. cv Sirosa) was compared on both a temporal and ontogenetic basis. As ontogeny and growth are interrelated, such a comparison allowed the growth and physiological responses to P level of the two species to be separated from responses due to the species being at different ontogenetic stages at the time of comparison. Plants were grown from seed through to anthesis under P deficient and P sufficient conditions in soil in a glasshouse. The ontogenies of Italian ryegrass and phalaris were similar, but the rate of development of Italian ryegrass was greater at both P rates. P deficiency resulted in arrested reproductive development in phalaris. At both P levels shoot, root and total biomass and net P uptake per plant by Italian ryegrass were greater than by phalaris when the two species were compared on a temporal basis, but when compared on an ontogenetic basis the two species were similar. There were some differences in the allocation of P between the acid-soluble P, lipid P, and residue P fractions, but biomass production was not determined by the efficiency of P utilization. The superior biomass production of Italian ryegrass on a temporal basis was due to its greater seed size and rate of ontogeny rather than differences in photosynthetic rate, unit leaf rate, leaf area ratio or shoot: root ratio. Similarly, the greater P uptake per plant of Italian ryegrass on a temporal basis was driven by its greater plant size and faster root extension rate rather than by P uptake per unit root length. The level of vesicular arbuscular mycorrhizal (VAM) infection in the roots of the two species was similar under P deficiency but greater in phalaris under P sufficient conditions. Overall, the different temporal responses to P of Italian ryegrass and phalaris were largely related to their different rates of ontogeny and the interrelationships between ontogeny and growth rate rather than to differences in their physiology in relation to P acquisition and utilization.

This study aimed to evaluate the efficiency of Mombasa grass in a silvopastoral system and submitted to different nitrogen fertilization rates and pasture management strategies. Treatments consisted of four nitrogen fertilizer doses (0.0, 125.0, 187.5, and 250.0 kg N ha-1 year-1) and three cutting heights (70, 90, and 105 cm). Plant agronomic and physiologic efficiencies, as well as nitrogen use and accumulation by plants, forage production, and increments of forage production, were verified for the application of nitrogen fertilizer. Nitrogen fertilization was only efficient for grasses cut at 105 cm, where 75% of the applied nitrogen at a dose of 125 kg ha-1 year-1 was recovered. Yet the grasses cut at 70 and 90 cm presented low results for nitrogen fertilization, showing plants with low physiological and agronomic efficiencies. For Mombasa grass in a silvopastoral system, nitrogen fertilization is more efficient when applied at lower doses and for plants cut at higher heights.

The objective of the present study was to analyze the influence of spray mixture volume and flight height on herbicide deposition in aerial applications on pastures. The experimental plots were arranged in a pasture area in the district of Porto Esperidião (Mato Grosso, Brazil). In all of the treatments, the applications contained the herbicides aminopyralid and fluroxypyr (Dominum) at the dose of 2.5 L c.p. ha-1, including the adjuvant mineral oil (Joint Oil) at the dose of 1.0 L and a tracer to determine the deposition by high-performance liquid chromatography (HPLC) (rhodamine at a concentration of 0.6%). The experiment consisted of nine treatments that comprised the combinations of three spray volumes (20, 30 and 50 L ha-1) and three flight heights (10, 30 and 40 m). The results showed that, on average, there was a tendency for larger deposits for the smallest flight heights, with a significant difference between the heights of 10 and 40 m. There was no significant difference among the deposits obtained with the different spray mixture volumes.

Há fortes evidências que as atividades humanas estão afetando o clima global por meio da produção de Gases de Efeito Estufa (GEE), dos quais o metano (CH4) tem elevado potencial de aquecimento. A fermentação entérica e o esterco dos ruminantes representam cerca de 30 a 40% do total das emissões antropogênicas de CH4. Este artigo traz um resumo de tecnologias existentes para reduzir as emissões de CH4 entéricas dos ruminantes, com ênfase à manipulação dietética e ruminal, à seleção/reprodução animal e à melhoria dos sistemas de produção. As diferenças na produção de CH4 entérica entre as espécies animais com base na anatomia do trato gastrointestinal, fisiologia digestiva, fermentação ruminal e nos hábitos de pastejo também são discutidas. A inibição da emissão de CH4 entérica é possível por meio do uso de ionóforos, ácidos orgânicos e óleos. Plantas alimentares contendo metabólitos secundários (taninos e saponinas i.e.) também podem reduzir a produção de CH4. O uso de animais reprodutores para melhorar a eficiência de conversão alimentar (menor consumo residual) pode contribuir com a redução da emissão total de CH4, além de reduzir a emissão por unidade de produto. Resultados utilizando o modelo IPCC nível II prevê que caprinos e vacas de elevada produção leiteira podem apresentar menor emissão de CH4 por unidade de produto em comparação com animais zebuínos e ovinos, enquanto os pequenos ruminantes (caprinos e ovinos) produzem menos CH4 por unidade de ganho de peso corporal (carne), em relação aos bovinos. A melhoria da qualidade das forragens e a implementação de práticas eficientes de uso das pastagens (sistema de pastejo e taxas de lotação) podem na maioria dos casos promover maior produção animal e incrementar a emissão absoluta de CH4, mas também reduzir a emissão de CH4 por unidade de produto animal. Palavras - chave: mudanças climáticas, gases do efeito estufa, bovinos, ovinos, caprinos, manipulação da dieta, manipulação ruminal, criação de animais, intensificação, modelagem. Estratégias para a Redução da Emissão de Metano por Ruminantes A B S T R A C T There is irrefutable evidence that human activities are affecting the global climate through the production of Green House Gases (GHG) of which methane (CH4) has a high warming potential. Enteric fermentation and manure from ruminants represent about 30 to 40% of the total anthropogenic CH4 emissions. This paper summarizes existing technologies to reduce enteric CH4 emissions in ruminants given emphasis to dietary and rumen manipulation, animal selection/ breeding and improvement of production systems. Differences in enteric CH4 production among animal species based on anatomy of the GI tract, digestive physiology, rumen fermentation and grazing habits are also discussed. Inhibition of enteric CH4 emission is possible through the use of ionophores, organic acids and oils. Feeding plants containing secondary metabolites (i.e. tannins and saponins) can reduce CH4 production. Breeding for improved feed conversion efficiency (lower residual feed intake) is likely to reduce total and per unit product CH4 emissions. Results using the IPCC Tier II model predict that goats and high producing dairy cattle can potentially produce less CH4 emissions per unit of milk than Cebu cattle or sheep, while small ruminants (goats and sheep) produce less CH4 per unit of live weight gain (meat) than cattle. The introduction of improved high quality forages and the implementation of efficient pasture utilization practices (grazing system and stocking rate) can result in most cases in improved animal production and in increased absolute CH4 emissions, but in reduced CH4 per unit of animal product. Keywords: climate change, greenhouse gases, cattle, sheep, goats, dietary manipulation, rumen manipulation, animal breeding, intensification, modelling

D’une famille française réfugiée à Genève au XVIe siècle, Jean Senebier fut successivement pasteur à Chancy, puis bibliothécaire à Genève. Charles Bonnet lui révéla sa veritable voie qui était naturaliste. II l'incita à traduire oeuvre expérimentale de Spallanzani. A son époque, la chimie des gaz faisait de grands progrès. Senebier eut le mérite d’appliquer cette chimie à Vetude des échanges gazeux de la plante. II apporta une contribution importante, tant au point de vue métkodologique qu’au point de vue physiologique.

The effects of shade on growth, biomass allocation patterns and photosynthetic response was examined for Rolandra fruticosa (L.) Kuntze, a common perennial weed shrub in cultivated pastures and agricultural areas of Brazilian Amazonia, for plants grown in full sunlight and those shaded to 30 % of full sunlight over a 34-d period. Specific leaf area and leaf area ratio were higher for shade plants during all the experimental period. Shade plants allocated significantly less biomass to root tissue than sun plants and relative growth rate was higher in sun plants. Sun leaves had significantly higher dark respiration and light saturated rates of photosynthesis than shade leaves. The apparent quantum efficiency was higher for shade leaves, while light compensation point was higher for sun leaves. These results are discussed in relation to their ecological and weed management implications.

ABSTRACT The species Tecoma stans, introduced to Brazil to be cultivated in urban centers as street trees, in parks and gardens, became important pasture weed. The mechanical control of this species is difficult because the plant presents a large number of sprouts when are mowed. Furthermore, the herbicides usually used in pasture have been shown to be inefficient to control it. This research evaluated the trumpet flower control effectiveness using a new herbicide (aminocyclopyrachlor) applied alone in increasing doses to the cut stump for the control of resprouts and mixed with metsulfuron-methyl in applications to intact plant leaves. The experiment with foliar applications used increasing doses of herbicides (aminocyclopyrachlor + metsulfuron-methyl). Applications of only aminocyclopyrachlor to the cut stump and foliar application of a herbicidal mixture (aminocyclopyrachlor + metsulfuron-methyl) were effective in the control of trumpet flower.

AbstractThe woolly-bear caterpillars of Grammia (= Apantesis) blakei (Grt.) are sometimes abundant on native range pastures in the short-grass region of southern Alberta and southwestern Saskatchewan and occasionally cause early season damage to adjacent cultivated crops. The larvae overwinter as mid- to late-instars in protected sites on the soil surface and begin feeding in the spring as soon as new plant growth is available. They are general feeders on both mono- and dicotyledonous plants. Larval densities exceeding 10/m2 were found at several locations from 1982 to 1985 and historical records indicate that densities sometimes exceed 50/m2. The moths are present in late May through June but are rarely observed or collected. In Canada the species is univoltine with a mid-larval summer diapause.

The potential for seed bank formation of two perennial weed species, Ipomoea asarifolia (Desr.) Roem. & Schult. (Convolvulaceae) and Stachytarpheta cayennensis (Rich.) M. Vahl (Verbenaceae), both common in Amazonia , was evaluated in a degraded pasture area in eastern Brazilian Amazonia . Seeds were enclosed in nylon mesh packets and placed at the soil surface or buried at 5 or 10 cm deep. The number of viable seeds was recorded at 6, 10, 14 and 18 months after burial. Results showed that S. cayennensis has the ability to form persistent soil seed bank, while I. asarifolia seeds do not build up in the soil seed bank. For S. cayennensis and, to some extent, for I. asarifolia, seed survival was highest at greater burial depths.

ABSTRACT Quinclorac is a systemic herbicide absorbed by germinating seeds, roots and leaves of seedlings. It is a selective compound for crops such as rice, canola, barley, corn, sorghum, and pasture. Quinclorac can be used to control various monocots and dicotyledonous weed species. The biochemical function of this herbicide in the plant has intrigued scientists for nearly four decades. The objectives of this review are to present evidence of three hypotheses on the biochemical functioning of quinclorac and to propose an integrative mode of action. The first theory on the mode of action of quinclorac is supported by evidence of inhibition of incorporation of C14-glucose into cellulose and hemicellulose, thus, affecting the cell wall synthesis. The second hypothesis suggests that quinclorac acts as an auxin in broadleaved weed species. In grass species, however, this herbicide appears to stimulate the activity of the 1-aminocyclopropane-1-carboxylate synthase enzyme and, subsequently, to increase the ethylene production; also, it seems to increase the cyanide acid content to phytotoxic levels. A third hypothesis to explain the harmful effect in some plant species is the formation of reactive oxygen species (ROS). Apparently, these processes are not mutually exclusive; therefore, an integrative theory for the action of quinclorac is suggested. It is theorized that the aforementioned biochemical activities are interconnected and can be the phytotoxic backbone to explain the herbicidal effect depending on the plant species and the plant growth stage, among other factors.

AbstractThe objectives of this study were to determine whether the spatial distribution of Melanoplus sanguinipes F., the most abundant species of grasshopper on rangeland in southern Idaho, varied annually in response to changing patterns of grazing and to investigate how vegetation affects the spatial distribution of low-density populations of M. sanguinipes at scales relevant to most rangeland-management activities. A lattice of 72 sites was established across nine pastures, covering approximately 5000 ha. At each site, densities of M. sanguinipes, percent canopy coverage by plant species, and percent forage utilization by livestock were estimated twice per year, in June when M. sanguinipes was in the nymphal stage and in August during the adult stage, for 4 years, 1991–1994. Spatial analyses of variance were used to evaluate the influence of grazing and vegetation type on densities of M. sanguinipes. In August of each year, densities of M. sanguinipes were lower on heavily grazed sites than on lightly grazed sites, except in 1993, when the opposite trend was observed. Above-normal precipitation in 1993 resulted in abundant growth of annual forbs and regrowth of grazed plants. The distribution of nymphs in June of 1993 and 1994 reflected the grazing patterns of the previous summer. Densities of M. sanguinipes were lower on crested wheatgrass habitats than on annual grasslands for every sampling period from June 1991 to June 1993, after which no differences were observed. We interpret the results to suggest that grazing effects on low-density populations of M. sanguinipes were contingent on weather conditions; under dry conditions, grazed habitats were less favorable to M. sanguinipes but, during relatively cool wet summers, grazing created conditions that were more favorable to M. sanguinipes.

Environments containing significant concentration of NaCl such as salt lakes harbor extremophiles microorganisms which have a great biotechnology interest. To explore the diversity of Bacteria in Chott Tinsilt (Algeria), an isolation program was performed. Water samples were collected from the saltern during the pre-salt harvesting phase. This Chott is high in salt (22.47% (w/v). Seven halophiles Bacteria were selected for further characterization. The isolated strains were able to grow optimally in media with 10–25% (w/v) total salts. Molecular identification of the isolates was performed by sequencing the 16S rRNA gene. It showed that these cultured isolates included members belonging to the Halomonas, Staphylococcus, Salinivibrio, Planococcus and Halobacillus genera with less than 98% of similarity with their closest phylogenetic relative. The halophilic bacterial isolates were also characterized for the production of biosurfactant and industrially important enzymes. Most isolates produced hydrolases and biosurfactants at high salt concentration. In fact, this is the first report on bacterial strains (A4 and B4) which were a good biosurfactant and coagulase producer at 20% and 25% ((w/v)) NaCl. In addition, the biosurfactant produced by the strain B4 at high salinity (25%) was also stable at high temperature (30-100°C) and high alkalinity (pH 11).Key word: Salt Lake, Bacteria, biosurfactant, Chott, halophiles, hydrolases, 16S rRNAINTRODUCTIONSaline lakes cover approximately 10% of the Earth’s surface area. The microbial populations of many hypersaline environments have already been studied in different geographical regions such as Great Salt Lake (USA), Dead Sea (Israel), Wadi Natrun Lake (Egypt), Lake Magadi (Kenya), Soda Lake (Antarctica) and Big Soda Lake and Mono Lake (California). Hypersaline regions differ from each other in terms of geographical location, salt concentration and chemical composition, which determine the nature of inhabitant microorganisms (Gupta et al., 2015). Then low taxonomic diversity is common to all these saline environments (Oren et al., 1993). Halophiles are found in nearly all major microbial clades, including prokaryotic (Bacteria and Archaea) and eukaryotic forms (DasSarma and Arora, 2001). They are classified as slight halophiles when they grow optimally at 0.2–0.85 M (2–5%) NaCl, as moderate halophiles when they grow at 0.85–3.4 M (5–20%) NaCl, and as extreme halophiles when they grow at 3.4–5.1 M (20–30%) NaCl. Hyper saline environments are inhabited by extremely halophilic and halotolerant microorganisms such as Halobacillus sp, Halobacterium sp., Haloarcula sp., Salinibacter ruber , Haloferax sp and Bacillus spp. (Solomon and Viswalingam, 2013). There is a tremendous demand for halophilic bacteria due to their biotechnological importance as sources of halophilic enzymes. Enzymes derived from halophiles are endowed with unique structural features and catalytic power to sustain the metabolic and physiological processes under high salt conditions. Some of these enzymes have been reported to be active and stable under more than one extreme condition (Karan and Khare, 2010). Applications are being considered in a range of industries such as food processing, washing, biosynthetic processes and environmental bioremediation. Halophilic proteases are widely used in the detergent and food industries (DasSarma and Arora, 2001). However, esterases and lipases have also been useful in laundry detergents for the removal of oil stains and are widely used as biocatalysts because of their ability to produce pure compounds. Likewise, amylases are used industrially in the first step of the production of high fructose corn syrup (hydrolysis of corn starch). They are also used in the textile industry in the de-sizing process and added to laundry detergents. Furthermore, for the environmental applications, the use of halophiles for bioremediation and biodegradation of various materials from industrial effluents to soil contaminants and accidental spills are being widely explored. In addition to enzymes, halophilic / halotolerants microorganisms living in saline environments, offer another potential applications in various fields of biotechnology like the production of biosurfactant. Biosurfactants are amphiphilic compounds synthesized from plants and microorganisms. They reduce surface tension and interfacial tension between individual molecules at the surface and interface respectively (Akbari et al., 2018). Comparing to the chemical surfactant, biosurfactant are promising alternative molecules due to their low toxicity, high biodegradability, environmental capability, mild production conditions, lower critical micelle concentration, higher selectivity, availability of resources and ability to function in wide ranges of pH, temperature and salinity (Rocha et al., 1992). They are used in various industries which include pharmaceuticals, petroleum, food, detergents, cosmetics, paints, paper products and water treatment (Akbari et al., 2018). The search for biosurfactants in extremophiles is particularly promising since these biomolecules can adapt and be stable in the harsh environments in which they are to be applied in biotechnology.OBJECTIVESEastern Algeria features numerous ecosystems including hypersaline environments, which are an important source of salt for food. The microbial diversity in Chott Tinsilt, a shallow Salt Lake with more than 200g/L salt concentration and a superficies of 2.154 Ha, has never yet been studied. The purpose of this research was to chemically analyse water samples collected from the Chott, isolate novel extremely or moderate halophilic Bacteria, and examine their phenotypic and phylogenetic characteristics with a view to screening for biosurfactants and enzymes of industrial interest.MATERIALS AND METHODSStudy area: The area is at 5 km of the Commune of Souk-Naâmane and 17 km in the South of the town of Aïn-Melila. This area skirts the trunk road 3 serving Constantine and Batna and the railway Constantine-Biskra. It is part the administrative jurisdiction of the Wilaya of Oum El Bouaghi. The Chott belongs to the wetlands of the High Plains of Constantine with a depth varying rather regularly without never exceeding 0.5 meter. Its length extends on 4 km with a width of 2.5 km (figure 1).Water samples and physico-chemical analysis: In February 2013, water samples were collected from various places at the Chott Tinsilt using Global Positioning System (GPS) coordinates of 35°53’14” N lat. and 06°28’44”E long. Samples were collected randomly in sterile polythene bags and transported immediately to the laboratory for isolation of halophilic microorganisms. All samples were treated within 24 h after collection. Temperature, pH and salinity were measured in situ using a multi-parameter probe (Hanna Instruments, Smithfield, RI, USA). The analytical methods used in this study to measure ions concentration (Ca2+, Mg2+, Fe2+, Na+, K+, Cl−, HCO3−, SO42−) were based on 4500-S-2 F standard methods described elsewhere (Association et al., 1920).Isolation of halophilic bacteria from water sample: The media (M1) used in the present study contain (g/L): 2.0 g of KCl, 100.0/200.0 g of NaCl, 1.0 g of MgSO4.7HO2, 3.0 g of Sodium Citrate, 0.36 g of MnCl2, 10.0 g of yeast extract and 15.0 g agar. The pH was adjusted to 8.0. Different dilutions of water samples were added to the above medium and incubated at 30°C during 2–7 days or more depending on growth. Appearance and growth of halophilic bacteria were monitored regularly. The growth was diluted 10 times and plated on complete medium agar (g/L): glucose 10.0; peptone 5.0; yeast extract 5.0; KH2PO4 5.0; agar 30.0; and NaCl 100.0/200.0. Resultant colonies were purified by repeated streaking on complete media agar. The pure cultures were preserved in 20% glycerol vials and stored at −80°C for long-term preservation.Biochemical characterisation of halophilic bacterial isolates: Bacterial isolates were studied for Gram’s reaction, cell morphology and pigmentation. Enzymatic assays (catalase, oxidase, nitrate reductase and urease), and assays for fermentation of lactose and mannitol were done as described by Smibert (1994).Optimization of growth conditions: Temperature, pH, and salt concentration were optimized for the growth of halophilic bacterial isolates. These growth parameters were studied quantitatively by growing the bacterial isolates in M1 medium with shaking at 200 rpm and measuring the cell density at 600 nm after 8 days of incubation. To study the effect of NaCl on the growth, bacterial isolates were inoculated on M1 medium supplemented with different concentration of NaCl: 1%-35% (w/v). The effect of pH on the growth of halophilic bacterial strains was studied by inoculating isolates on above described growth media containing NaCl and adjusted to acidic pH of 5 and 6 by using 1N HCl and alkaline pH of 8, 9, 10, 11 and 12 using 5N NaOH. The effect of temperature was studied by culturing the bacterial isolates in M1 medium at different temperatures of incubation (4°C–55°C).Screening of halophilic bacteria for hydrolytic enzymes: Hydrolase producing bacteria among the isolates were screened by plate assay on starch, tributyrin, gelatin and DNA agar plates respectively for amylase, lipase, protease and DNAse activities. Amylolytic activity of the cultures was screened on starch nutrient agar plates containing g/L: starch 10.0; peptone 5.0; yeast extract 3.0; agar 30.0; NaCl 100.0/250.0. The pH was 7.0. After incubation at 30 ºC for 7 days, the zone of clearance was determined by flooding the plates with iodine solution. The potential amylase producers were selected based on ratio of zone of clearance diameter to colony diameter. Lipase activity of the cultures was screened on tributyrin nutrient agar plates containing 1% (v/v) of tributyrin. Isolates that showed clear zones of tributyrin hydrolysis were identified as lipase producing bacteria. Proteolytic activity of the isolates was similarly screened on gelatin nutrient agar plates containing 10.0 g/L of gelatin. The isolates showing zones of gelatin clearance upon treatment with acidic mercuric chloride were selected and designated as protease producing bacteria. The presence of DNAse activity on plates was determined on DNAse test agar (BBL) containing 10%-25% (w/v) total salt. After incubation for 7days, the plates were flooded with 1N HCl solution. Clear halos around the colonies indicated DNAse activity (Jeffries et al., 1957).Milk clotting activity (coagulase activity) of the isolates was also determined following the procedure described (Berridge, 1952). Skim milk powder was reconstituted in 10 mM aqueous CaCl2 (pH 6.5) to a final concentration of 0.12 kg/L. Enzyme extracts were added at a rate of 0.1 mL per mL of milk. The coagulation point was determined by manual rotating of the test tube periodically, at short time intervals, and checking for visible clot formation.Screening of halophilic bacteria for biosurfactant production. Oil spread Assay: The Petridis base was filled with 50 mL of distilled water. On the water surface, 20μL of diesel and 10μl of culture were added respectively. The culture was introduced at different spots on the diesel, which is coated on the water surface. The occurrence of a clear zone was an indicator of positive result (Morikawa et al., 2000). The diameter of the oil expelling circles was measured by slide caliber (with a degree of accuracy of 0.02 mm).Surface tension and emulsification index (E24): Isolates were cultivated at 30 °C for 7 days on the enrichment medium containing 10-25% NaCl and diesel oil as the sole carbon source. The medium was centrifuged (7000 rpm for 20 min) and the surface tension of the cell-free culture broth was measured with a TS90000 surface tensiometer (Nima, Coventry, England) as a qualitative indicator of biosurfactant production. The culture broth was collected with a Pasteur pipette to remove the non-emulsified hydrocarbons. The emulsifying capacity was evaluated by an emulsification index (E24). The E24 of culture samples was determined by adding 2 mL of diesel oil to the same amount of culture, mixed for 2 min with a vortex, and allowed to stand for 24 h. E24 index is defined as the percentage of height of emulsified layer (mm) divided by the total height of the liquid column (mm).Biosurfactant stability studies : After growth on diesel oil as sole source of carbone, cultures supernatant obtained after centrifugation at 6,000 rpm for 15 min were considered as the source of crude biosurfactant. Its stability was determined by subjecting the culture supernatant to various temperature ranges (30, 40, 50, 60, 70, 80 and 100 °C) for 30 min then cooled to room temperature. Similarly, the effect of different pH (2–11) on the activity of the biosurfactant was tested. The activity of the biosurfactant was investigated by measuring the emulsification index (El-Sersy, 2012).Molecular identification of potential strains. DNA extraction and PCR amplification of 16S rDNA: Total cellular DNA was extracted from strains and purified as described by Sambrook et al. (1989). DNA was purified using Geneclean® Turbo (Q-BIO gene, Carlsbad, CA, USA) before use as a template in polymerase chain reaction (PCR) amplification. For the 16S rDNA gene sequence, the purified DNA was amplified using a universal primer set, forward primer (27f; 5′-AGA GTT TGA TCM TGG CTC AG) and a reverse primer (1492r; 5′-TAC GGY TAC CTT GTT ACG ACT T) (Lane, 1991). Agarose gel electrophoresis confirmed the amplification product as a 1400-bp DNA fragment.16S rDNA sequencing and Phylogenic analysis: Amplicons generated using primer pair 27f-1492r was sequenced using an automatic sequencer system at Macrogene Company (Seoul, Korea). The sequences were compared with those of the NCBI BLAST GenBank nucleotide sequence databases. Phylogenetic trees were constructed by the neighbor-joining method using MEGA version 5.05 software (Tamura et al., 2011). Bootstrap resembling analysis for 1,000 replicates was performed to estimate the confidence of tree topologies.Nucleotide sequence accession numbers: The nucleotide sequences reported in this work have been deposited in the EMBL Nucleotide Sequence Database. The accession numbers are represented in table 5.Statistics: All experiments were conducted in triplicates. Results were evaluated for statistical significance using ANOVA.RESULTSPhysico-chemical parameters of the collected water samples: The physicochemical properties of the collected water samples are reported in table 1. At the time of sampling, the temperature was 10.6°C and pH 7.89. The salinity of the sample, as determined in situ, was 224.70 g/L (22,47% (w/v)). Chemical analysis of water sample indicated that Na +and Cl- were the most abundant ions (table 1). SO4-2 and Mg+2 was present in much smaller amounts compared to Na +and Cl- concentration. Low levels of calcium, potassium and bicarbonate were also detected, often at less than 1 g/L.Characterization of isolates. Morphological and biochemical characteristic feature of halophilic bacterial isolates: Among 52 strains isolated from water of Chott Tinsilt, seven distinct bacteria (A1, A2, A3, A4, B1, B4 and B5) were chosen for further characterization (table 2). The colour of the isolates varied from beige, pale yellow, yellowish and orange. The bacterial isolates A1, A2, A4, B1 and B5 were rod shaped and gram negative (except B5), whereas A3 and B4 were cocci and gram positive. All strains were oxidase and catalase positive except for B1. Nitrate reductase and urease activities were observed in all the bacterial isolates, except B4. All the bacterial isolates were negative for H2S formation. B5 was the only strain positive for mannitol fermentation (table 2).We isolated halophilic bacteria on growth medium with NaCl supplementation at pH 7 and temperature of 30°C. We studied the effect of NaCl, temperature and pH on the growth of bacterial isolates. All the isolates exhibited growth only in the presence of NaCl indicating that these strains are halophilic. The optimum growth of isolates A3 and B1 was observed in the presence of 10% NaCl, whereas it was 15% NaCl for A1, A2 and B5. A4 and B4 showed optimum growth in the presence of 20% and 25% NaCl respectively. A4, B4 and B5 strains can tolerate up to 35% NaCl.The isolate B1 showed growth in medium supplemented with 10% NaCl and pH range of 7–10. The optimum pH for the growth B1 was 9 and they did not show any detectable growth at or below pH 6 (table 2), which indicates the alkaliphilic nature of B1 isolate. The bacterial isolates A1, A2 and A4 exhibited growth in the range of pH 6–10, while A3 and B4 did not show any growth at pH greater than 8. The optimum pH for growth of all strains (except B1) was pH 7.0 (table 2). These results indicate that A1, A2, A3, A4, B4 and B5 are neutrophilic in nature. All the bacterial isolates exhibited optimal growth at 30°C and no detectable growth at 55°C. Also, detectable growth of isolates A1, A2 and A4 was observed at 4°C. However, none of the bacterial strains could grow below 4°C and above 50°C (table 2).Screening of the halophilic enzymes: To characterize the diversity of halophiles able to produce hydrolytic enzymes among the population of microorganisms inhabiting the hypersaline habitats of East Algeria (Chott Tinsilt), a screening was performed. As described in Materials and Methods, samples were plated on solid media containing 10%-25% (w/v) of total salts and different substrates for the detection of amylase, protease, lipase and DNAse activities. However, coagulase activity was determined in liquid medium using milk as substrate (figure 3). Distributions of hydrolytic activity among the isolates are summarized in table 4.From the seven bacterial isolates, four strains A1, A2, A4 and B5 showed combined hydrolytic activities. They were positive for gelatinase, lipase and coagulase. A3 strain showed gelatinase and lipase activities. DNAse activities were detected with A1, A4, B1 and B5 isolates. B4 presented lipase and coagulase activity. Surprisingly, no amylase activity was detected among all the isolates.Screening for biosurfactant producing isolates: Oil spread assay: The results showed that all the strains could produce notable (>4 cm diameter) oil expelling circles (ranging from 4.11 cm to 4.67 cm). The average diameter for strain B5 was 4.67 cm, significantly (P < 0.05) higher than for the other strains.Surface tension and emulsification index (E24): The assimilation of hydrocarbons as the sole sources of carbon by the isolate strains led to the production of biosurfactants indicated by the emulsification index and the lowering of the surface tension of cell-free supernatant. Based on rapid growth on media containing diesel oil as sole carbon source, the seven isolates were tested for biosurfactant production and emulsification activity. The obtained values of the surface tension measurements as well as the emulsification index (E24) are shown in table 3. The highest reduction of surface tension was achieved with B5 and A3 isolates with values of 25.3 mN m−1 and 28.1 mN m−1 respectively. The emulsifying capacity evaluated by the E24 emulsification index was highest in the culture of isolate B4 (78%), B5 (77%) and A3 (76%) as shown in table 3 and figure 2. These emulsions were stable even after 4 months. The bacteria with emulsification indices higher than 50 % and/or reduction in the surface tension (under 30 mN/m) have been defined as potential biosurfactant producers. Based on surface tension and the E24 index results, isolates B5, B4, A3 and A4 are the best candidates for biosurfactant production. It is important to note that, strains B4 and A4 produce biosurfactant in medium containing respectively 25% and 20% (w/v) NaCl.Stability of biosurfactant activities: The applicability of biosurfactants in several biotechnological fields depends on their stability at different environmental conditions (temperatures, pH and NaCl). For this study, the strain B4 appear very interesting (It can produce biosurfactant at 25 % NaCl) and was choosen for futher analysis for biosurfactant stability. The effects of temperature and pH on the biosurfactant production by the strain B4 are shown in figure 4.biosurfactant in medium containing respectively 25% and 20% (w/v) NaCl.Stability of biosurfactant activities: The applicability of biosurfactants in several biotechnological fields depends on their stability at different environmental conditions (temperatures, pH and NaCl). For this study, the strain B4 appear very interesting (It can produce biosurfactant at 25 % NaCl) and was chosen for further analysis for biosurfactant stability. The effects of temperature and pH on the biosurfactant production by the strain B4 are shown in figure 4. The biosurfactant produced by this strain was shown to be thermostable giving an E-24 Index value greater than 78% (figure 4A). Heating of the biosurfactant to 100 °C caused no significant effect on the biosurfactant performance. Therefore, the surface activity of the crude biosurfactant supernatant remained relatively stable to pH changes between pH 6 and 11. At pH 11, the value of E24 showed almost 76% activity, whereas below pH 6 the activity was decreased up to 40% (figure 4A). The decreases of the emulsification activity by decreasing the pH value from basic to an acidic region; may be due to partial precipitation of the biosurfactant. This result indicated that biosurfactant produced by strain B4 show higher stability at alkaline than in acidic conditions.Molecular identification and phylogenies of potential isolates: To identify halophilic bacterial isolates, the 16S rDNA gene was amplified using gene-specific primers. A PCR product of ≈ 1.3 kb was detected in all the seven isolates. The 16S rDNA amplicons of each bacterial isolate was sequenced on both strands using 27F and 1492R primers. The complete nucleotide sequence of 1336,1374, 1377,1313, 1305,1308 and 1273 bp sequences were obtained from A1, A2, A3, A4, B1, B4 and B5 isolates respectively, and subjected to BLAST analysis. The 16S rDNA sequence analysis showed that the isolated strains belong to the genera Halomonas, Staphylococcus, Salinivibrio, Planococcus and Halobacillus as shown in table 5. The halophilic isolates A2 and A4 showed 97% similarity with the Halomonas variabilis strain GSP3 (accession no. AY505527) and the Halomonas sp. M59 (accession no. AM229319), respectively. As for A1, it showed 96% similarity with the Halomonas venusta strain GSP24 (accession no. AY553074). B1 and B4 showed for their part 96% similarity with the Salinivibrio costicola subsp. alcaliphilus strain 18AG DSM4743 (accession no. NR_042255) and the Planococcus citreus (accession no. JX122551), respectively. The bacterial isolate B5 showed 98% sequence similarity with the Halobacillus trueperi (accession no. HG931926), As for A3, it showed only 95% similarity with the Staphylococcus arlettae (accession no. KR047785). The 16S rDNA nucleotide sequences of all the seven halophilic bacterial strains have been submitted to the NCBI GenBank database under the accession number presented in table 5. The phylogenetic association of the isolates is shown in figure 5.DICUSSIONThe physicochemical properties of the collected water samples indicated that this water was relatively neutral (pH 7.89) similar to the Dead Sea and the Great Salt Lake (USA) and in contrast to the more basic lakes such as Lake Wadi Natrun (Egypt) (pH 11) and El Golea Salt Lake (Algeria) (pH 9). The salinity of the sample was 224.70 g/L (22,47% (w/v). This range of salinity (20-30%) for Chott Tinsilt is comparable to a number of well characterized hypersaline ecosystems including both natural and man-made habitats, such as the Great Salt Lake (USA) and solar salterns of Puerto Rico. Thus, Chott Tinsilt is a hypersaline environment, i.e. environments with salt concentrations well above that of seawater. Chemical analysis of water sample indicated that Na +and Cl- were the most abundant ions, as in most hypersaline ecosystems (with some exceptions such as the Dead Sea). These chemical water characteristics were consistent with the previously reported data in other hypersaline ecosystems (DasSarma and Arora, 2001; Oren, 2002; Hacěne et al., 2004). Among 52 strains isolated from this Chott, seven distinct bacteria (A1, A2, A3, A4, B1, B4 and B5) were chosen for phenotypique, genotypique and phylogenetique characterization.The 16S rDNA sequence analysis showed that the isolated strains belong to the genera Halomonas, Staphylococcus, Salinivibrio, Planococcus and Halobacillus. Genera obtained in the present study are commonly occurring in various saline habitats across the globe. Staphylococci have the ability to grow in a wide range of salt concentrations (Graham and Wilkinson, 1992; Morikawa et al., 2009; Roohi et al., 2014). For example, in Pakistan, Staphylococcus strains were isolated from various salt samples during the study conducted by Roohi et al. (2014) and these results agreed with previous reports. Halomonas, halophilic and/or halotolerant Gram-negative bacteria are typically found in saline environments (Kim et al., 2013). The presence of Planococcus and Halobacillus has been reported in studies about hypersaline lakes; like La Sal del Rey (USA) (Phillips et al., 2012) and Great Salt Lake (Spring et al., 1996), respectively. The Salinivibrio costicola was a representative model for studies on osmoregulatory and other physiological mechanisms of moderately halophilic bacteria (Oren, 2006).However, it is interesting to note that all strains shared less than 98.7% identity (the usual species cut-off proposed by Yarza et al. (2014) with their closest phylogenetic relative, suggesting that they could be considered as new species. Phenotypic, genetic and phylogenetic analyses have been suggested for the complete identification of these strains. Theses bacterial strains were tested for the production of industrially important enzymes (Amylase, protease, lipase, DNAse and coagulase). These isolates are good candidates as sources of novel enzymes with biotechnological potential as they can be used in different industrial processes at high salt concentration (up to 25% NaCl for B4). Prominent amylase, lipase, protease and DNAase activities have been reported from different hypersaline environments across the globe; e.g., Spain (Sánchez‐Porro et al., 2003), Iran (Rohban et al., 2009), Tunisia (Baati et al., 2010) and India (Gupta et al., 2016). However, to the best of our knowledge, the coagulase activity has never been detected in extreme halophilic bacteria. Isolation and characterization of crude enzymes (especially coagulase) to investigate their properties and stability are in progress.The finding of novel enzymes with optimal activities at various ranges of salt concentrations is of great importance. Besides being intrinsically stable and active at high salt concentrations, halophilic and halotolerant enzymes offer great opportunities in biotechnological applications, such as environmental bioremediation (marine, oilfiel) and food processing. The bacterial isolates were also characterized for production of biosurfactants by oil-spread assay, measurement of surface tension and emulsification index (E24). There are few reports on biosurfactant producers in hypersaline environments and in recent years, there has been a greater increase in interest and importance in halophilic bacteria for biomolecules (Donio et al., 2013; Sarafin et al., 2014). Halophiles, which have a unique lipid composition, may have an important role to play as surface-active agents. The archae bacterial ether-linked phytanyl membrane lipid of the extremely halophilic bacteria has been shown to have surfactant properties (Post and Collins, 1982). Yakimov et al. (1995) reported the production of biosurfactant by a halotolerant Bacillus licheniformis strain BAS 50 which was able to produce a lipopeptide surfactant when cultured at salinities up to 13% NaCl. From solar salt, Halomonas sp. BS4 and Kocuria marina BS-15 were found to be able to produce biosurfactant when cultured at salinities of 8% and 10% NaCl respectively (Donio et al., 2013; Sarafin et al., 2014). In the present work, strains B4 and A4 produce biosurfactant in medium containing respectively 25% and 20% NaCl. To our knowledge, this is the first report on biosurfactant production by bacteria under such salt concentration. Biosurfactants have a wide variety of industrial and environmental applications (Akbari et al., 2018) but their applicability depends on their stability at different environmental conditions. The strain B4 which can produce biosurfactant at 25% NaCl showed good stability in alkaline pH and at a temperature range of 30°C-100°C. Due to the enormous utilization of biosurfactant in detergent manufacture the choice of alkaline biosurfactant is researched (Elazzazy et al., 2015). On the other hand, the interesting finding was the thermostability of the produced biosurfactant even after heat treatment (100°C for 30 min) which suggests the use of this biosurfactant in industries where heating is of a paramount importance (Khopade et al., 2012). To date, more attention has been focused on biosurfactant producing bacteria under extreme conditions for industrial and commercial usefulness. In fact, the biosurfactant produce by strain B4 have promising usefulness in pharmaceutical, cosmetics and food industries and for bioremediation in marine environment and Microbial enhanced oil recovery (MEOR) where the salinity, temperature and pH are high.CONCLUSIONThis is the first study on the culturable halophilic bacteria community inhabiting Chott Tinsilt in Eastern Algeria. Different genera of halotolerant bacteria with different phylogeneticaly characteristics have been isolated from this Chott. Culturing of bacteria and their molecular analysis provides an opportunity to have a wide range of cultured microorganisms from extreme habitats like hypersaline environments. Enzymes produced by halophilic bacteria show interesting properties like their ability to remain functional in extreme conditions, such as high temperatures, wide range of pH, and high salt concentrations. These enzymes have great economical potential in industrial, agricultural, chemical, pharmaceutical, and biotechnological applications. Thus, the halophiles isolated from Chott Tinsilt offer an important potential for application in microbial and enzyme biotechnology. In addition, these halo bacterial biosurfactants producers isolated from this Chott will help to develop more valuable eco-friendly products to the pharmacological and food industries and will be usefulness for bioremediation in marine environment and petroleum industry.ACKNOWLEDGMENTSOur thanks to Professor Abdelhamid Zoubir for proofreading the English composition of the present paper.CONFLICT OF INTERESTThe authors declare that they have no conflict of interest.Akbari, S., N. H. Abdurahman, R. M. Yunus, F. Fayaz and O. R. Alara, 2018. Biosurfactants—a new frontier for social and environmental safety: A mini review. Biotechnology research innovation, 2(1): 81-90.Association, A. P. H., A. W. W. Association, W. P. C. 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Schleifer, W. B. Whitman, J. Euzéby, R. Amann and R. Rosselló-Móra, 2014. Uniting the classification of cultured and uncultured bacteria and archaea using 16s rRNA gene sequences. Nature reviews microbiology, 12(9): 635-645

ABSTRACT: The great diversity of plant species in pastures of the Brazilian Cerrado with distinct ecophysiological characteristics indicates the possibility of finding, in the cellular content of this flora, chemical compounds with potential for use in agriculture and human and animal health. Three steps are necessary to prove this hypothesis: phytochemical prospecting, characterization of secondary metabolites, and studies on the biological activities of these metabolites present in these plants. The chemical profile of secondary metabolites present in five species of the Brazilian Cerrado (Davilla elliptica, Remijia ferruginea, Luehea paniculata, Anacardium occidentale, and Acosmium dasycarpum) was traced in this research. These plant species were collected in pasture areas of Felício dos Santos, Minas Gerais. The samples were dried and submitted to two types of extract (ethanolic and hexanic). By using specific chemical reactions, the presence of coumarins, triterpenes/steroids, and anthracenosides was observed in the hexanic extracts, while the presence of alkaloids, triterpenes/steroids, flavonoids, tannins, reducing compounds, and anthocyanins was observed in the ethanolic extracts. The species presented diverse classes of compounds. However, triterpenes/steroids, tannins, reducing compounds and anthocyanins were found in all species, being the use of the solvent ethanol the most efficient in extracting the compounds. The species D. elliptica presented the highest number of classes of compounds. The widespread application in folk medicine justifies further studies on the biological activity of different metabolites in agriculture and health areas.

ABSTRACT: This research aimed at evaluating the effect of increasing coexistence periods with weeds on the nutritional components of the forage grass Brachiaria brizantha cv. Marandú under pasture renewal conditions. The experiment was arranged in randomized blocks with four replications, and treatments were represented by eight coexistence periods, namely: 0, 15, 30, 45, 60, 75, 90 and 120 days after seedling emergence. A phytosociologic evaluation was performed on the weed community at the end of the coexistence periods; later, weeds were eliminated by an herbicide formulated with aminopyralid+2,4-D (40+320 g L-1) at 2.5 L ha-1, applied in post-emergence. The fodder plants were evaluated at the end of the experimental period as for leaf-stem ratio, volumetric density of green leaf, volumetric density of total dry matter and main nutritional components, including: crude protein, neutral detergent fiber, ether extract, organic matter and mineral matter, and from this, the total carbohydrate content was calculated. The presence of weeds altered the pasture structure, since it affected negatively the leaf/stem ratio and the volumetric leaf density of B. brizantha. The coexistence also reduced the nutritional components of B. brizantha and determined that control measures should be adopted before reaching 30 days of coexistence between fodder plants and weeds.

ABSTRACT: The grazing behavior practice by bovines can be positive or negatively influenced by the pasture structure where the animal is inserted. Several factors determine the pasture structure and the presence of weeds is considered one of the most important. This study aimed to assess the effect of species with and without stiff structures over the grazing behavior of bovines in pasture areas. The experimental design was a split block design with four replications, in which treatments were arranged in a 4 x 3 factorial design: four weed species (Zanthoxylum rhoifolium, Cnidoscolus urens, Dasyphyllum brasiliensis, and Luehea divaricata) associated with three proximity strips of weeds: 0-50, 50-100, and 100-150 cm in relation to the main stem of the studied weed. Forage intake by animals was measured by determining the real forage offers at 0, 3, 6, 9, and 15 days after the beginning of animal grazing (DAP). The presence of noxious shrubs influenced bovine grazing behavior. The influence on the intake is most evident in the presence of plants that promote animal discomfort. The species Z. rhoifolium, C. urens, and D. brasiliensis were the weeds with the greatest influence on feed access among the species that have stiff structures. The negative influence on grazing behavior is higher in the proximity strip closest to the plant, i.e. 0-50 cm from the main stem of the weed.

ABSTRACT: In pastures, the incidence of weeds reduces the productivity and quality of forage. The identification of the weed species in pastures is fundamental to choose the renewal method. Thus, the objective of this work was to perform phytosociology before and after the renewal of a Brachiaria decumbens pasture, with the implantation of agrosilvopastoral systems. Eighteen different crop arrangements for pasture renewal were evaluated. The renewal systems were by eucalyptus integration (at 12 x 2 m or 12 x 3 m spacings) with maize, Brachiaria brizantha cv. Marandu (palisade grass) and/or Macrotyloma axillare (perennial horsegram), or monoculture and intercropping of palisade grass and perennial horsegram, as well as the evaluation of the application or not of the herbicide bentazon, at the recommended dose for maize crops (0.72 kg ha-1). Relative frequency, relative density, relative abundance, relative dominance, coverage value index, importance value index, dry matter and similarity index were evaluated. In the first survey, before the pasture renewal, 23 plant species were identified. After the implantation of agrosilvopastoral systems, the species Sida cordifolia, Lantana camara and B. decumbens were the only occurring ones before and after the renewal of the pasture with agrosilvopastoral systems. The use of palisade grass and the application of the herbicide were efficient in controlling weeds. In systems that contained palisade grass and perennial horsegram, the latter was not found in the survey conducted one year after the implantation.

ABSTRACT: Pantanal plain has large extensions of land formed by many types of landscapes suitable to extensive livestock system. Open grasslands and lowlands are formed by forage such as grasses and forbs widely consumed by cattle. However, climatic and human factors can promote the dissemination of invasive shrub species such as canjiqueira (Byrsonima cydoniifolia A. Juss) making it necessary, in many cases, the human intervention to control the invasion. The present work had as objective to determine the economic threshold level to control canjiqueira through two methods: tractor driven blade and link chains pulled by tractor in different invasion levels plants per hectare. Economic Threshold proved to be a tool for efficient and effective management for making decisions of when and how to do the control of canjiqueira in natural pastures of Pantanal of Nhecolândia.

ABSTRACT: Corn and forage intercropping system has been commonly used for pasture establishment or renewal. In addition, the use of reduced doses of herbicides selective to corn has facilitated coexistence by reducing competitiveness against forage species. This study aimed to evaluate the effects of mesotrione underdoses in a corn and Urochloa brizantha intercropping on weed population dynamics, corn yield, and forage performance and nutritional quality. For this purpose, five mesotrione doses were tested (0.0, 9.6, 19.2, 38.4, and 57.6 g ha-1) in corn and forage intercropping and monocultures. Individual density and weed dry matter were influenced by mesotrione doses, intercropping, and precipitation distribution over the evaluated period in the intercropping and pasture. In intercropping, Alternanthera tenella and Commelina benghalensis were the most important species because they could not be controlled by the herbicide. After intercropping, A. tenella and C. benghalensis presented higher relative importance (RI) in the formed pasture during the rainy season, while Conyza bonariensis and A. tenella presented the highest RI during the dry season. Corn grain yield was not affected by the presence of grass, but forage yield was affected by the competition with corn. Mesotrione doses did not affect the yield and nutritional quality of the forage. The effects on mineral matter, crude protein, and acid detergent fiber in the forage were due to the coexistence with corn when compared to the control grown in monoculture.

ABSTRACT: The phenological stage, post-harvest processing and quantity of material can interfere in the allelopathic activity of extracts. The objective of this study is to test the allelopathic effect of aqueous extracts of the leaves of Annoni grass (Eragrostis plana), an invader of natural pastures. Twelve extracts were prepared by combining the phenological stage of the plants when harvesting the leaves (vegetative; flowering), post-harvest processing before extraction (fresh; dry at 40 oC), and the amount of plant material (5, 15, and 25 g 100 mL-1 distilled water). Two bioassays were conducted in a germination chamber, with evaluation of germination and growth of seedlings of lettuce (Lactuca sativa) and white clover (Trifolium repens). The extracts with higher allelopathic activity were further developed with 15 and 25 g of dried leaves and harvested in plants at the vegetative stage. Germination was attributed to being the greatest contribution to the divergence between the extracts. White clover was more sensitive to extracts, for which 50% to 67% of the extracts were effectively allelopathic (≥50% inhibition); in lettuce, between 8% and 58% of the extracts exhibited this potential. The phenological stage was the factor with a greater individual effect on the allelopathic activity on lettuce and white clover seedlings, and on lettuce germination. The post-harvest processing responded by the greater variation on germination and germination speed index of white clover. The sensitivity of the white clover to extracts of Annoni grass suggests a compromise of its establishment in pastures with the presence of the invader.

ABSTRACT: This research aimed to study weed interference relationship on morphogenesis, yield, and greenhouse gas production potential of Urochloa ruziziensis under pasture renovation conditions. The experimental design was a randomized block design with four replications. Treatments consisted of seven coexistence periods: 0, 15, 30, 45, 60, 75, and 90 days after emergence (DAE). The following morphological parameters were analyzed: number of tillers, number of leaves, photosynthetically active leaf blade biomass, fresh stem biomass, dead material biomass, and total dry biomass. In vitro analyses of methane (CH4) and carbon dioxide (CO2) production were also carried out. The results were submitted to analysis of variance by the F-test, and the test of means was carried out by the Scott-Knott test at 5%. Biomass, CH4, and CO2 production of U. ruziziensis were modified with only 15 days of weed coexistence, with an effect even higher from 45 DAE. Therefore, weeds interfere with all morphological parameters, yield, and greenhouse gas production in the pasture establishment with the forage grass U. ruziziensis.

ABSTRACT: The aim of this study was to evaluate the establishment of the pasture of Panicum maximum cv. Aries in an environmentally protected area under levels of interspecific interaction with the weed community. The experiment started after sowing P. maximum, and it was carried out in a randomized block design with four replications and the following factors: 3 light interception levels, 2 nitrogen rates, and 7 weekly sampling dates along pasture establishment (3x2x7). The light interception treatments, determined by photosynthetically active radiation, were given by mowing weeds over the canopy of P. maximum at 40% and 70% light interception, and no mowing (uncontrolled growth of weeds and P. maximum). Topdressing application of nitrogen in the form of urea was performed or not (0 or 200 kg N ha-1). The plant community was evaluated by number of species, dry matter accumulation, and density, and phytosociological indices were determined. The relative importance and dry matter accumulation of P. maximum were greater after nitrogen fertilization, which favored the species against weed competition. In contrast, there was lower weed density without nitrogen fertilization. Mowing weeds at 40% of light interception enabled P. maximum to accumulate more dry matter, while there was no difference between mowing weeds at 70% light interception and growing the forage crop freely with weeds. Reducing weed light interception as well as nitrogen fertilization, and consequently reducing the competitive ability of the weed, favored the establishment of P. maximum in diversified agro-ecosystems.

ABSTRACT: Two lipophilic extracts and atomaric acid (1), an isolated natural product, were obtained from the marine brown alga Stypopodium zonale (Dictyotaceae) to identify and characterize their potential inhibitory effects on the seed germination, radicle elongation, and hypocotyl development of the weeds Mimosa pudica and Senna obtusifolia. The extracts were prepared with hexane and dichloromethane, and atomaric acid (1) was isolated from hexane extract by way of conventional chromatographic methods. During a 15 days period, germination bioassays were performed at 25 oC with a 12 h photoperiod, whereas radicle elongation and hypocotyl development were assayed at 25 oC with a 24 h photoperiod. After, Petri dishes 9.0 cm in diameter were coated with qualitative filter paper, 25 seeds were placed in a germination chamber, while six pregerminated seeds were placed in the Petri dish for 2-3 days. After 10 days, radicle and hypocotyl extension were measured; and the inhibitory potential of the extracts was assessed at 10 ppm and that of the atomaric acid at 5, 10, 15, and 20 ppm. In both M. pudica and S. obtusifolia, dichloromethane extract achieved the greatest rates of inhibition during seed germination (34% and 22%, respectively), radical germination (38% and 30%, respectively), and hypocotyl development (29% and 22%, respectively). At a concentration of 20 ppm, atomaric acid (1) also demonstrated reduced inhibitory potential, with mean values of 58.67% for M. pudica and 48.67% for S. obtusifolia.

ABSTRACT: The Brazilian native weed assa-peixe (Vernonanthura brasiliana) is considered one of the main invasive plant species of pasture areas in the Amazon. It is thus necessary to know their biology and their behavior in the environment, and to enhance the development of management strategies, minimizing the use of herbicides. The objective of this study was to test the allelopathic effect on the germination and growth of the bioindicator species Lactuca sativa from leaf and root extracts obtained from fresh and dry V. brasiliana. The experiment consisted of six treatments with concentrations of 0, 12.5, 25, 50, 100, and 200 mg mL-1, using four replicates per treatment. The evaluated parameters were: germination percentage, first count, germination speed index, and shoot and root length. The aqueous extract of fresh V. brasiliana roots caused an allelopathic effect on all evaluated parameters, while the dry root extract only affected the germination speed index. Root growth inhibition and growth stimulation of lettuce seedlings were verified when submitted to dry and fresh root extracts. Extracts of V. brasiliana caused an allelopathic effect on the species Lactuca sativa L., where the aqueous extracts of dry leaves and fresh roots were the most influential.

ABSTRACT: The addition of adjuvants to herbicide solutions is aimed at preserving or enhancing the biological effect of treatment. However, it is commonly performed without knowledge of the physicochemical interactions between products. This study aimed to assess the effects of different addition sequences of the herbicide aminopyralid + fluroxypyr and adjuvants in the preparation of phytosanitary spray solutions on the surface tension and contact angle. Two experiments were carried out with herbicide doses of 1 and 2 L ha-1 associated with the adjuvants mineral oil (MO), silicone-polyether copolymer (SIL), and a mixture of phosphatidylcholine (lectin) and propionic acid (LEC), all at a proportion of 0.3% v v-1. The application rate was 150 L ha-1. Surface tension was measured by the pendant droplet method. Contact angle was measured on the adaxial and abaxial surfaces of leaves of the pasture weed Senna obtusifolia and parafilm. Preparation sequence did not change the contact angle on any of the analyzed surfaces at a dose of 1 L ha-1 of herbicide. For the dose of 2 L ha-1, the adjuvants SIL and LEC showed a higher spreading when previously added to the herbicide. MO resulted in a higher spreading when added after the herbicide, with higher surface coverage. Therefore, the preparation sequence influences the dispersion of phytosanitary spray solutions on targets.

ABSTRACT: Razor grass (Paspalum virgatum L) is one weed that has gain importance because of its high seed production capability, mainly when pasture fields need to be renewed. Understanding seedling emergence behavior is crucial to devise strategies to manage this weed. This trial was carried out to evaluate the emergence pattern of razor grass seedlings on the basis of seed depth in the soil profile. The experiment was conducted in a green-house in a completely randomized design, with six replications. The treatments performed in a 11 x 3 factorial arrangement: eleven seeding depths (surface, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0 and 10.0 cm) by three ecotypes (Rondonópolis, Redenção and Cacoal, Brazil). The effect of origin and seed depth was measured by number of emerged seedlings, emergence speed index, relative emergence frequency, mean emergence time and emergence synchrony of seedlings at 35 days after planting. The seeds of ecotype Rondonópolis had higher vigor than those of ecotype Redenção, which had higher vigor that the ones of ecotype Cacoal. The highest seedling emergence percentage, uniformity and speed were found when the seeds were located at the soil surface, regardless of seed origin. There was no seedling emergence when the seeds of ecotypes Rondonópolis and Cacoal were located at a depth equal to or higher than 8.0 cm; and of ecotype Redenção when they were at a depth equal or higher than 7.0 cm.