Search results for: direct inoculation

Authors: Simonida S. Djuric, Timea I. Hajnal Jafari, Dragana R. Stamenov

Abstract:

The use of plant growth promoting fungi (PGPF) has significantly increased in the last decade mostly due to their multi-level properties, and their expected success as biofertilizers in agriculture. Beneficial fungi with broad-host range undergo long-term interactions with a large variety of plants thereby playing a significant role in managed ecosystems and in the adaptation of crops to global climate changes. Trichoderma spp. are promising fungi toward the development of sustainable agriculture. The aim of our experiment was to investigate the effect of seed inoculation of sunflower, maize, soybean, paprika, melon, and watermelon seeds with Trichoderma sp. on early seed germination energy and initial growth of the plant. The seed inoculation with Trichoderma sp. increased the seedling emergence from 7, 85% in melon to 156,70% in watermelon. The inoculation had the best effect on initial growth of maize shoot (+23,80%) and soybean root (+106,30%). The different response of seed and young plants on Trichoderma sp. inoculation implicate the need for future investigations of successful inoculation systems and modes of their integration in sustainable agriculture production systems.

Keywords: initial growth, inoculation, seedling, Trichoderma sp.

Procedia PDF Downloads 206

Authors: Ahmed Elgharably, Nivien Nafady

Abstract:

Arbuscular mycorrhizal fungi (AMF) and plant growth promoting microbes (PGPM) can promote plant growth under saline conditions. This study investigated how AMF and PGPM affected the growth and grain yield of wheat at different soil salinity levels (0, 75 and 150 mM NaCl). AMF colonization percentage, grain yield and dry weights and lengths of shoot and root, N, P K, Na, malondialdehyde, chlorophyll and proline contents and shoot relative permeability were determined. Salinity reduced NPK uptake and malondialdehyde and chlorophyll contents, and increased shoot Na concentration, relative permeability, and proline content, and thus declined plant growth. PGPM inoculation enhanced AMF colonization, P uptake, and K/Na ratio, but alone had no significant effect on plant growth and grain yield. AMF inoculation significantly enhanced NPK uptake, increased chlorophyll content and decreased shoot relative permeability, proline and Na contents, and thus promoted the plant growth. The inoculation of PGPM significantly enhanced the positive effects of AMF in controlling Na uptake and in increasing chlorophyll and NPK contents. Compared to AMF inoculation alone, dual inoculation with AMF and PGPM resulted in approximately 10, 25 and 25% higher grain yield at 0, 75 and 150 mM NaCl, respectively. The results provide that PGPM inoculation can maximize the effects of AMF inoculation in alleviating the deleterious effects of NaCl salts on wheat growth.

Keywords: mycorrhizal fungi, salinity, sodium, wheat

Procedia PDF Downloads 148

Authors: M. S. Alatas, H. D. Umucalilar

Abstract:

SARA is a common and serious metabolic disorder in early lactation in dairy cattle and in finishing beef cattle, caused by diets with high inclusion of cereal grain. This experiment was performed to determine the efficacy of Megasphaera elsdenii, a major lactate-utilizing bacterium in prevention/treatment of SARA in vivo. In vivo experimentation, it was used eight ruminally cannulated rams and it was applied the rapid adaptation with the mixture of grain based on wheat (%80 wheat, %20 barley) and barley (%80 barley, %20 wheat). During the systematic adaptation, it was followed the probability of SARA formation by being measured the rumen pH with two hours intervals after and before feeding. After being evaluated the data, it was determined the ruminal pH ranged from 5,2-5,6 on the condition of feeding with 60 percentage of grain mixture based on barley and wheat, that assured the definite form of subacute acidosis. In four days SARA period, M. elsdenii (1010 cfu ml-1) was inoculated during the first two days. During the SARA period, it was observed the decrease of feed intake with M. elsdenii inoculation. Inoculation of M. elsdenii was caused to differentiation of rumen pH (P < 0,0001), while it was found the pH level approximately 5,55 in animals applied the inoculation, it was 5,63 pH in other animals. It was observed that total VFA with the bacterium inoculation tended to change in terms of grain feed (P < 0,07). It increased with the effect of total VFA inoculation in barley based diet, but it was more stabilized in wheat based diet. Bacterium inoculation increased the ratio of propionic acid (18,33%-21,38%) but it caused to decrease the butyric acid, and acetic/propionic acid. During the rapid adaptation, the concentration of lactic acid in the rumen liquid increased depending upon grain level (P<0,0001). On the other hand bacterium inoculation did not have an effect on concentration of lactic acid. M. elsdenii inoculation did not affect ruminal ammonia concentration. In the group that did not apply inoculation, the level of ruminal ammonia concentration was higher than the others applied inoculation. M. elsdenii inoculation did not changed protozoa count in barley-based diet whereas it decreased in wheat-based diet. In the period of SARA, it was observed that the level of blood glucose, lactate and hematocrit increased greatly after inoculation (P < 0,0001). When it is generally evaluated, it is seen that M. elsdenii inoculation has not a positive impact on rumen parameters. Therefore, to reveal the full impact of the inoculation with different strains, feedstuffs and animal groups, further research is required.

Keywords: In vivo, Subactute ruminal acidosis, Megasphaera elsdenii, Rumen fermentation

Procedia PDF Downloads 608

Authors: Spartak Makovskyi, Volodymir Klochykhin, Valery Zakharchenko, Konstantyn Balushok, Eduard Tsyvirko, Anatoly Shalomeyev

Abstract:

A highly efficient, cost-effective grain refinement technique for ML5 magnesium alloy with a commercially pure carbon nanopowder has been proposed. An experimental casting of testing specimens with incremental additions of a carbon nanopowder (0.001 - 0.1 wt.% ) was performed. It has been found that the carbon nanoparticle inoculation of the alloy structure is efficient in a narrow concentration range. The additions of 0.005-0.01 wt. % the grain refiner in the alloy resulted in a maximum increase of ductility properties (appr. Twofold) and improved tensile strength. However, further expansion of the grain refiner content led to the deterioration of the alloy's mechanical properties. In particular, the introduction of 0.1 wt.% of the nanocarbon and more caused internal defects in the metal. The carbon nanoparticle inoculation is a promising way of improving the properties of the Mg-Al-Zn alloys for critical lightweight aerospace applications on an industrial scale.

Keywords: carbon nanopowder, inoculation, melt, tensile strength

Procedia PDF Downloads 171

Authors: Waad Al Saleemi, Badriya Al Adawi, Zaaima Al Jabri, Sahim Al Ghafri, Jalila Al Hadhramia

Abstract:

Objectives: Using standard lab methods, a positive blood culture requires a minimum of two days (two occasions of overnight incubation) to obtain a final identification (ID) and antimicrobial susceptibility results (AST) report. In this study, we aimed to evaluate the accuracy and precision of identification and antimicrobial susceptibility testing of an alternative method (direct method) that will reduce the turnaround time by 24 hours. This method involves the direct inoculation of positive blood culture broths into the Phoenix system using serum separation tubes (SST). Method: This prospective study included monomicrobial-positive blood cultures obtained from January 2022 to May 2023 in SQUH. Blood cultures containing a mixture of organisms, fungi, or anaerobic organisms were excluded from this study. The result of the new “direct method” under study was compared with the current “standard method” used in the lab. The accuracy and precision were evaluated for the ID and AST using Clinical and Laboratory Standards Institute (CLSI) recommendations. The categorical agreement, essential agreement, and the rates of very major errors (VME), major errors (ME), and minor errors (MIE) for both gram-negative and gram-positive bacteria were calculated. Passing criteria were set according to CLSI. Result: The results of ID and AST were available for a total of 158 isolates. Of 77 isolates of gram-negative bacteria, 71 (92%) were correctly identified at the species level. Of 70 isolates of gram-positive bacteria, 47(67%) isolates were correctly identified. For gram-negative bacteria, the essential agreement of the direct method was ≥92% when compared to the standard method, while the categorical agreement was ≥91% for all tested antibiotics. The precision of ID and AST were noted to be 100% for all tested isolates. For gram-positive bacteria, the essential agreement was >93%, while the categorical agreement was >92% for all tested antibiotics except moxifloxacin. Many antibiotics were noted to have an unacceptable higher rate of very major errors including penicillin, cotrimoxazole, clindamycin, ciprofloxacin, and moxifloxacin. However, no error was observed in the results of vancomycin, linezolid, and daptomycin. Conclusion: The direct method of ID and AST for positive blood cultures using SST is reliable for gram negative bacteria. It will significantly decrease the turnaround time and will facilitate antimicrobial stewardship.

Keywords: bloodstream infection, oman, direct ast, blood culture, rapid identification, antimicrobial susceptibility, phoenix, direct inoculation

Procedia PDF Downloads 18

Authors: Said Cheto, Khawla Oukaltouma, Imane Chamkhi, Ammar Ibn Yasser, Bouchra Benmrid, Ahmed Qaddoury, Lamfeddal Kouisni, Joerg Geistlinger, Youssef Zeroual, Adnane Bargaz, Cherki Ghoulam

Abstract:

Our study aimed to assess, the role of inoculation of faba bean/wheat intercrops with selected rhizobacteria consortia gathering one rhizobia and two phosphate solubilizing bacteria “PSB” to alleviate the effects of combined water deficit and P limitation on Faba bean/ wheat intercrops versus monocrops under greenhouse conditions. One Vicia faba L variety (Aguadulce “Ag”), and one Triticum durum L. variety (Karim “K”) were grown as sole crops or intercrop in pots containing sterilized substrate (sand: peat 4:1v/v) added either with rock phosphate (RP) as the alone P source (P limitation) or with KH₂PO₄ in nutrient solution (P sufficient control). Plant inoculation was done using rhizobacterial consortia composed; C1(Rhizobium laguerreae, Kocuria sp, and Pseudomonas sp) and C2 (R. laguerreae, Rahnella sp, and Kocuria sp). Two weeks after inoculation, the plants were submitted to water deficit consisting of 40% of substrate water holding Capacity (WHC) versus 80% WHC for well-watered plants. At the flowering stage, the trial was assessed, and the results showed that inoculation with both consortia (C1 and C2) improved faba bean biomass in terms of shoots, roots, and nodules compared to inoculation with rhizobia alone, particularly C2 improved these parametres by 19.03, 78.99, and 72.73%, respectively. Leaf relative water content decreased under combined stress, particularly in response to C1 with a significant improvement of this parameter in wheat intercrops. For faba bean under P limitation, inoculation with C2 increased stomatal conductance (gs) by 35.73% compared to plants inoculated with rhizobia alone. Furthermore, the same inoculum C2 improved membrane stability by 44,33% versus 16,16% for C1 compared to inoculation with rhizobia alone under P deficit. For sole cropped faba bean plants, inoculation with both consortia improved N accumulation compared to inoculation with rhizobia alone with an increase of 70.75% under P limitation. Moreover, under the combined stress, intercropping inoculation with C2 improved plant biomass and N content (112.98%) in wheat plants, compared to the sole crop. Our finding revealed that consortium C2 might offer an agronomic advantage under water and P deficit and could be used as inoculum for enhancing faba bean and wheat production under both monocropping and intercropping systems.

Keywords: drought, phosphorus, intercropping, PSB, rhizobia, vicia faba, Triticum durum

Procedia PDF Downloads 41

Authors: F. Z. Aliyat, B. Ben Messaoud, L. Nassiri, E. Bouiamrine, J. Ibijbijen

Abstract:

The present study aims to assess the biological nitrogen fixation in the soybean tested in different Moroccan soils combined with the rhizobial inoculation. These effects were evaluated by the plant growth mainly by the aerial biomass production, total nitrogen content and the proportion of the nitrogen fixed. This assessment clearly shows that the inoculation with bacteria increases the growth of soybean. Five different soils and a control (peat) were used. The rhizobial inoculation was performed by applying the peat that contained a mixture of 2 strains Sinorhizobium fredii HH103 and Bradyrhizobium. The biomass, the total nitrogen content and the proportion of nitrogen fixed were evaluated under different treatments. The essay was realized at the greenhouse the Faculty of Sciences, Moulay Ismail University. The soybean has shown a great response for the parameters assessed. Moreover, the best response was reported by the inoculated plants compared to non- inoculated and to the absolute control. Finally, good production and the best biological nitrogen fixation present an important ecological technology to improve the sustainable production of soybean and to ensure the increase of the fertility of soils.

Keywords: biological nitrogen fixation, inoculation, rhizobium, soybean

Procedia PDF Downloads 140

Authors: Timea I. Hajnal-Jafari, Simonida S. Đurić, Dragana R. Stamenov

Abstract:

Microbial inoculants from the group of symbiotic-nitrogen-fixing rhizobia are well known and widely used in production of legumes. On the other hand, nonsymbiotic plant growth promoting rhizobacteria (PGPR) are not commonly used in practice. The objective of this study was to examine the effects of soybean inoculation with symbiotic and nonsymbiotic bacteria on plant growth and seed yield of soybean. Microbiological activity in rhizospheric soil was also determined. The experiment was set up using a randomized block system in filed conditions with the following treatments: control-no inoculation; treatment 1-Bradyrhizobium japonicum; treatment 2-Azotobacter sp.; treatment 3-Bacillus sp..In the flowering stage of growth (FS) the number of nodules per plant (NPP), root length (RL), plant height (PH) and weight (PW) were measured. The number of pod per plant (PPP), number of seeds per pod (SPP) and seed weight per plant (SWP) were recorded at the end of vegetation period (EV). Microbiological analyses of soil included the determination of total number of bacteria (TNB), number of fungi (FNG), actinomycetes (ACT) and azotobacters (AZB) as well as the activity of the dehydrogenase enzyme (DHA). The results showed that bacterial inoculation led to the formation of root nodules regardless of the treatments with statistically no significant difference. Strong nodulation was also present in control treatment. RL and PH were positively influenced by inoculation with Azotobacter sp. and Bacillus sp., respectively. Statistical analyses of the number of PPP, SPP, and SWP showed no significant differences among investigated treatments. High average number of microorganisms were determined in all treatments. Most abundant were TNB (log No 8,010) and ACT (log No 6,055) than FNG and AZB with log No 4,867 and log No 4,025, respectively. The highest DHA activity was measured in the FS of soybean in treatment 3. The application of nonsymbiotic bacteria in soybean production can alleviate initial plant growth and help the plant to better overcome different stress conditions caused by abiotic and biotic factors.

Keywords: bacteria, inoculation, soybean, microbial activity

Procedia PDF Downloads 118

Authors: Oqba Basal, Andras Szabo

Abstract:

Using mineral fertilization is increasing worldwide, as it is claimed to be majorly responsible for achieving high yields; however, the negative impacts of mineral fertilization on soil and environment are becoming more obvious, with alternative methods being more necessary and applicable, especially with the current climatic changes which have imposed serious abiotic stresses, such as drought. An experiment was made during 2017 growing season in Debrecen, Hungary to investigate the effects of inoculation and N fertilization on the seed yield and protein concentration of the soybean (Glycine max (L.) Merr.) cultivar (Panonia Kincse) under three different irrigation regimes: severe drought stress (SD), moderate drought stress (MD) and control with no drought stress (ND). Three N fertilizer rates were applied: no N fertilizer (0 N), 35 kg ha⁻¹ of N fertilizer (35 N) and 105 kg ha⁻¹ of N fertilizer (105 N). Half of the seeds in each treatment was inoculated with Bradyrhizobium japonicum inoculant, and the other half was not inoculated. The results showed significant differences in the seed yield associated with inoculation, irrigation and the interaction between them, whereas there were no significant differences in the seed yield associated with fertilization alone or in interaction with inoculation or irrigation or both. When seeds were inoculated, yield was increased when (35 N) was applied compared to (0 N) but not significantly; however, the high rate of N fertilizer (105 N) reduced the yield to a level even less than (0 N). When seeds were not inoculated, the highest rate of N increased the yield the most compared to the other two N fertilizer rates whenever the drought was present (moderate or severe). Under severe drought stress, inoculation was positively and significantly correlated with yield; however, adding N fertilizer increased the yield of uninoculated plants compared to the inoculated ones, regardless of the rate of N fertilizer. Protein concentration in the seeds was significantly affected by irrigation and by fertilization, but not by inoculation. Protein concentration increased as the N fertilization rate increased, regardless of the inoculation or irrigation treatments; moreover, increasing the N rate reduced the correlation coefficient of protein concentration with the irrigation. It was concluded that adding N fertilizer is not always recommended, especially when seeds are inoculated before being sown; however, it is very important under severe drought stress to sustain yield. Enhanced protein concentrations could be achieved by applying N fertilization, whether the seeds were pre-inoculated or not.

Keywords: drought stress, N fertilization, protein concentration, soybean

Procedia PDF Downloads 124

Authors: Mubashar Iqbal, Iqra Anjum, Muhammad Dildar Gogi, Muhammad Jalal Arif

Abstract:

The present study was carried to screen out the effective entomopathogenic fungi (EPF) inoculation method in maize and to evaluate pathogenicity and oviposition-choice in C. partellus. Three entomopathogenic fungi (EPF) formulations Pacer® (Metarhizium anisopliae), Racer® (Beauveria bassiana) and Meailkil® (Verticillium lecanii) were evaluated at three concentrations (5000, 10000 and 20000 ppm) for their endophytism in maize and pathogenicity in C. partellus. The stock solution of the highest concentration (20,000 ppm) was prepared and next lower from stock solution. In the first experiment, three EPF was inoculated in maize plant by four methods, i.e., leaf-inoculation (LI), whorl-inoculation (WI), shoot-inoculation (SI) and root-inoculation (RI). Leaf-discs and stem-cutting were sampled in all four inoculation methods and placed on fungus growth media in Petri dishes. In the second experiment, pathogenicity, pupal formation, adult emergence, sex ratio, oviposition-choice, and growth index of C. partellus were calculated. The leaves and stem of the inoculated plants were given to the counted number of larvae of C. Partellus. The mortality of larvae was recorded on daily basis till the pupation. The result shows that maximum percent mortality (86.67%) was recorded at high concentration (20000ppm) of Beauveria bassiana by leaf inoculation method. For oviposition choice bioassay, the newly emerged adults were fed on diet (water, honey and yeast in 9:1:1) for 48 hours. One pair of C. Partellus were aspirated from the rearing cages and were detained in large test tube plugged with diet soaked cotton. A set of four plants for each treatment were prepared and randomized inside the large oviposition chamber. The test tubes were opened and fitted in the hole made in the wall of oviposition chamber in front of each treatment. The oviposition chamber was placed in a completely dark laboratory to eliminate the effect of light on moth’s behavior. The plants were removed from the oviposition chamber after the death of adults. The number of eggs deposited on the plant was counted. The results of 2nd experiment revealed that in all EPF and inoculation methods, the fecundity, egg fertility and growth index of C. partellus decreased with the increase in concentration being significantly higher at low concentration (5000ppm) and lower at higher concentration (20000ppm). Application of B. bassiana demonstrated that minimum fecundity (126.83), egg fertility (119.52) and growth index (15%) in C. partellus followed by M. anisopliae with fecundity (135.93), egg fertility (132.29) and growth index (17.50%) while V. lecanii show higher values of fecundity (137.37), egg fertility (1135.42) and growth index (20%). Overall leaf inoculation method showed least fecundity (123.89) with egg fertility (115.36) and growth index (14%) followed by whorl, shoot inoculation method and root inoculation method show higher values of fecundity, egg fertility and growth index.

Keywords: Beauveria bassiana, Chilo partellus, entomopathoganic, Metarhizium anisopliae, Verticillium lecanii

Procedia PDF Downloads 100

Authors: N. S. Aggangan, B. Dell, P. Jeffries

Abstract:

Two moderately nickel-tolerant isolates of Pisolithus were compared with a non-Ni tolerant isolate for the ability to increase the growth of Eucalyptus urophylla seedlings in the presence of nickel (Ni) in pots in a glasshouse. Seedlings, either inoculated with mycorrhizal fungi or uninoculated, were transplanted into pots containing 3 kg steam-pasteurized yellow sand amended with five concentrations of nickel (0, 6, 12, 24 and 48 mg Ni kg-1 soil). Within a day after transplanting, all seedlings subjected to Ni rates greater than 12 mg Ni kg-1 showed symptoms of wilting and all died within two weeks. At lower nickel concentrations, inoculation with all 3 Pisolithus strains increased rates of seedling survival after 12 weeks. Inoculation with all 3 isolates Pisolithus significantly increased the growth of plants in Ni-free soils between 2 to 4 fold dependent on isolate. However, seedlings growing in soils containing 12 mg Ni kg-1 grew poorly, mycorrhizal development was inhibited and no beneficial effects of inoculation were noted. In contrast, in soils containing 6mg Ni kg-1, inoculated seedlings did not show the reduced root growth and severe toxicity symptoms (chlorosis on young leaves and shoot tips) of uninoculated seedlings. Only the Ni-tolerant Pisolithus strains conferred a significant growth benefit compared to non-inoculated controls, and plants inoculated with one of these strains grew twice the size as those inoculated with the other Ni-tolerant strain. Inorganic plant analysis revealed that inoculation increased plant growth through improved P uptake but did not prevent Ni uptake. However, toxicity may have been minimized by dilution due to an increase in plant biomass. The results suggest that only one of the Ni-tolerant strains of Pisolithus has the potential to improve the growth and survival of E. urophylla seedlings in serpentine soils in the Philippines.

Keywords: ectomycorrhizas, Eucalyptus urophylla, nickel tolerance, pisolithus

Procedia PDF Downloads 275

Authors: A. Djoudi, R. Djibaou, H. A. Reguieg Yssaad

Abstract:

Our study focuses on the study of a bacteria belonging to Pseudomonas solubilizing tricalcium phosphate. They were isolated from rhizosphere of a variety of broad bean grown in salt-affected soils (electrical conductivity between 4 and 8 mmhos/cm) of the irrigated perimeter of Mina in northwestern Algeria. Isolates which have advantageous results in the calcium phosphate solubilization index test were subjected to identification using API20 then used to re-inoculate the same soil in pots experimentation to assess the effects of inoculation on the growth of the broad bean (Vicia faba). Based on the results obtained from the in-vitro tests, two isolates P5 and P8 showed a significant effect on the solubilization of tricalcium phosphate with an index I estimated at 314% and 283% sequentially. According to the results of in-vivo tests, the inoculation of the soil with P5 and P8 were significantly and positively influencing the growth in biometric parameters of the broad bean. Inoculation with strain P5 has promoted the growth of the broad bean in stem height, stem fresh weight and stem dry weight of 108.59%, 115.28%, 104.33%, respectively. Inoculation with strain P8 has fostered the growth of the broad bean stem fresh weight of 112.47%. The effect of Pseudomonas on the development of Vicia faba is considered as an interesting process by which PGPR can increase biological production and crop protection.

Keywords: Pseudomonas, Vicia faba, promoting of plant growth, solubilization tricalcium phosphate

Procedia PDF Downloads 299

Authors: Emmanuel Ekpa, Adaji Andrew, Queen Opaluwa, Isreal Daraobong

Abstract:

Three (3) indigenous bacteria species (Bacillus spp, Acinectobacter spp and Moraxella spp) previously isolated from contaminated soil of some auto mechanic workshops were used for bioremediation studies on some irrigation water used at Sarkin-noma Fadama farms located in Lokoja Kogi State, Nigeria. This was done in order to investigate their bioremediation potentials using a simple pour plate method. The physicochemical parameters and heavy metal analysis (using AAS iCE 3000) of the irrigation water were performed before and after inoculation of the isolated organisms. Nitrate and phosphate concentration were found to be 10.56mg/L and 12.63mg/L prior to inoculation while iron and zinc were 0.9569mg/L and 0.2245mg/L respectively. Other physicochemical parameters were also observed to be high prior to inoculation. After the bioremediation test (inoculation with the isolated organisms), a nitrate and phosphate content of 2.53mg/L and 2.61mg/L were recorded respectively, iron and zinc gave 0.1694mg/L and 0.0174mg/L concentrations while other physicochemical parameters measured were also found to be lower in their respective values. The implication of this present study is that a number of carefully isolated indigenous bacteria species are capable of reducing the amount of heavy metal concentrations in water. Also, non-metallic contaminants like nitrate and phosphate are susceptible to bioremediation in the presence of such efficient system.

Keywords: bioremediation, heavy metals, physicochemical parameters, Bacillus spp, Acinectobacter spp and Moraxella spp, AAS, spectrometer 3000

Procedia PDF Downloads 301

Authors: Rupa Rani, Vipin Kumar

Abstract:

Endosulfan, an organochlorine pesticide is of environmental concern due to its apparent persistence and toxicity. It has been reported as contaminants in soil, air, and water and is bioaccumulated and magnified in ecosystems. The combined use of microorganisms and plants has great potential for remediating soil contaminated with organic compounds such as pesticides. The objective of this study was to evaluate whether the bacterial inoculation influences plant growth promotion, endosulfan degradation in soil and endosulfan accumulation in different plant parts. Lycopersicon esculentum L. (Tomato) was grown in endosulfan spiked soil and inoculated with endosulfan tolerant bacterial strains. Endosulfan residues from different parts of plants and soil were extracted and estimated by using gas chromatograph equipped with 63Ni electron capture detector (GC-ECD). The inoculation of bacterial strains into the soil with plants showed a beneficial effect on endosulfan degradation and plant biomass production. Maximum endosulfan (90%) degradation was observed after 120 days of bacterial inoculation in the soil. Furthermore, there was significantly less endosulfan accumulation in roots and shoots of bacterial strains inoculated plants as compared to uninoculated plants. The results show the effectiveness of inoculated endosulfan tolerant bacterial strains to increase the remediation of endosulfan contaminated soil.

Keywords: organochlorine pesticides, endosulfan, degradation, plant-bacteria partnerships

Procedia PDF Downloads 122

Authors: B. D. Nkosi, T. F. Mutavhatsindi, J. J. Baloyi, R. Meeske, T. M. Langa, I. M. M. Malebana, M. D. Motiang

Abstract:

Potato hash (PH), a by-product from food production industry, contains 188.4 g dry matter (DM)/kg and 3.4 g water soluble carbohydrate (WSC)/kg DM, and was mixed with wheat bran (70:30 as is basis) to provide 352 g DM/kg and 315 g WSC/kg DM. The materials were ensiled with or without silage additives in 1.5L anaerobic jars (3 jars/treatment) that were kept at 25-280 C for 3 months. Four types of silages were produced which were: control (no additive, denoted as T1), celluclast enzyme (denoted as T2), emsilage bacterial inoculant (denoted as T3) and silosolve bacterial inoculant (denoted as T4). Three jars per treatment were opened after 3 months of ensiling for the determination of nutritive values, fermentation characteristics and aerobic stability. Aerobic stability was done by exposing silage samples to air for 5 days. The addition of enzyme (T2) was reduced (P<0.05) silage pH, fiber fractions (NDF and ADF) while increasing (P < 0.05) residual WSC and lactic acid (LA) production, compared to other treatments. Silage produced had pH of < 4.0, indications of well-preserved silage. Bacterial inoculation (T3 and T4) improved (P < 0.05) aerobic stability of the silage, as indicated by increased number of hours and lower CO2 production, compared to other treatments. However, the aerobic stability of silage was worsen (P < 0.05) with the addition of an enzyme (T2). Further work to elucidate these effects on nutrient digestion and growth performance on ruminants fed the silage is needed.

Keywords: by-products, digestibility, feeds, inoculation, ruminants, silage

Procedia PDF Downloads 408

Authors: Nour Elhouda Abed, Bedj Mimi, Wahid Slimani, Mourad Atif, Abdelhakim Ouzzane, Hocine Irekti, Abdelkader Bekki

Abstract:

The most frequent system of cereal production in Algeria is fallow-wheat. This is an extensive system that meets only the half needs some cereals and fodder demand. Resorption of fallow has become a strategic necessity to ensure food security in response to the instability of supply and the persistence of higher food prices on the world market. Despite several attempts to replace the fallow by crop cultures, choosing the best crop remains. Today, the agronomic and economic interests of legumes are demonstrated. However, their crop culture remains marginalized because of the weakness and instability of their performance. In the context of improving legumes and cereals crops as well as fallow resorption, we undertook to test, in the field, the effect of rhizobial inoculation of Phaseolus vulgaris in association with Zea Mays. We firstly studied the genetic diversity of rhizobial strains that nodulate P.vulgaris isolated from fifteen (15) different regions. ARDRA had shown 18 different genetic profiles. Symbiotic characterization highlighted a strain that highly significantly improved the fresh and dry weight of the host plant, in comparison to the negative control (un-inoculated) and the positive control (inoculated with the reference strain CIAT 899). In the field, the selected strain increased significantly the growth and yield of P.vulgaris and Zea Mays comparing to the non-inoculated control. However, the mix inoculation (selected strain+ Ciat 899) had not given the best parameters showing, thus, no synergy between the strains. These results indicate the replacing fallow by a crop legume in intercropping with cereals crops.

Keywords: fallow, intercropping, inoculation, legumes-cereals

Procedia PDF Downloads 334

Authors: Steven Mavhungu, Didier Nyembwe, Daniel Sekotlong

Abstract:

The steering knuckle is an integral component of the suspension and stability control system of modern vehicles. Good mechanical properties with an emphasis on the fatigue properties are essential for this component as it is subjected to cyclical load of significant magnitude during service. These properties are a function of the microstructure achieved in the component during the various manufacturing processes including forging and casting. The strut mount of the knuckle is required to meet specified microstructure and mechanical properties. However, in line with the recent trend of stringent quality requirements of cast components, Original Equipment Manufacturers (OEMs) have had to extend the specifications to other sections of the knuckle. This paper evaluates the effect of cored wire inoculation on the microstructure and mechanical properties of the steering arm of a typical spheroidal cast iron component. The investigation shows that the use of a cored wire having higher rare earth content formulation could possibly lead to a homogeneous matrix containing consistent graphite nodule morphology. However, this was found not to be the condition for better mechanical properties along the knuckle arm in line with required specifications. The findings in this paper contribute to a better understanding of steering knuckle properties to allow its production for safer automobile applications.

Keywords: inoculation, magnesium cored wire, spheroidal graphie, steering knuckle

Procedia PDF Downloads 194

Authors: John K. Mensah, Mary A. Sword Sayer, Ryan L. Nadel, George Matusick, Zhaofei Fan, Lori G. Eckhardt

Abstract:

Leptographium terebrantis is an opportunistic root pathogen commonly associated with loblolly pine (Pinus taeda L.) stands that are undergoing a loss of vigor in the southeastern US. In order to understand the relationship between L. terebrantis inoculum density and host physiology, an artificial inoculation study was conducted in a five-year-old naturally regenerated loblolly pine stand over a 24 week period in a completely randomized design. L. terebrantis caused sapwood occlusions that increased in severity as inoculum density increased. The occlusions significantly reduced water transport through the stem but did not interfere with fascicle-level stomatal conductance or induce moisture stress in the saplings. The resilience of stomatal conductance among pathogen-infested saplings is attributed to the growth and hydraulic function of new sapwood that developed after artificial inoculation. Results demonstrate that faster-growing families of loblolly pine may be capable of tolerating the vascular root disease when the formation of new sapwood is supported by sustained crown health.

Keywords: hydraulic conductance, inoculum density, Leptographium terebrantis, Pinus taeda, sapwood occlusion

Procedia PDF Downloads 293

Authors: Oqba Basal, András Szabó

Abstract:

Legume crops are able to fix atmospheric nitrogen by the symbiotic relation with specific bacteria, which allows the use of the mineral nitrogen-fertilizer to be reduced, or even excluded, resulting in more profit for the farmers and less pollution for the environment. Soybean (Glycine max (L.) Merrill) is one of the most important legumes with its high content of both protein and oil. However, it is recommended to combine the two nitrogen sources under stress conditions in order to overcome its negative effects. Drought stress is one of the most important abiotic stresses that increasingly limits soybean yields. A precise rate of mineral nitrogen under drought conditions is not confirmed, as it depends on many factors; soybean yield-potential and soil-nitrogen content to name a few. An experiment was conducted during 2017 growing season in Debrecen, Hungary to investigate the effects of nitrogen source on the physiology and the yield of the soybean cultivar 'Boglár'. Three N-fertilizer rates including no N-fertilizer (0 N), 35 kg ha^-1 of N-fertilizer (35 N) and 105 kg ha^-1 of N-fertilizer (105 N) were applied under three different irrigation regimes; severe drought stress (SD), moderate drought stress (MD) and control with no drought stress (ND). Half of the seeds in each treatment were pre-inoculated with Bradyrhizobium japonicum inoculant. The overall results showed significant differences associated with fertilization and irrigation, but not with inoculation. Increasing N rate was mostly accompanied with increased chlorophyll content and leaf area index, whereas it positively affected the plant height only when the drought was waived off. Plant height was the lowest under severe drought, regardless of inoculation and N-fertilizer application and rate. Inoculation increased the yield when there was no drought, and a low rate of N-fertilizer increased the yield furthermore; however, the high rate of N-fertilizer decreased the yield to a level even less than the inoculated control. On the other hand, the yield of non-inoculated plants increased as the N-fertilizer rate increased. Under drought conditions, adding N-fertilizer increased the yield of the non-inoculated plants compared to their inoculated counterparts; moreover, the high rate of N-fertilizer resulted in the best yield. Regardless of inoculation, the mean yield of the three fertilization rates was better when the water amount increased. It was concluded that applying N-fertilizer to provide the nitrogen needed by soybean plants, with the absence of N₂-fixation process, is very important. Moreover, adding relatively high rate of N-fertilizer is very important under severe drought stress to alleviate the drought negative effects. Further research to recommend the best N-fertilizer rate to inoculated soybean under drought stress conditions should be executed.

Keywords: drought stress, inoculation, N-fertilizer, soybean physiology, yield

Procedia PDF Downloads 116

Authors: Inuwa Abdu Ibrahim

Abstract:

The research assessed representative democracy as an administrative system in Nigeria, by highlighting the failure of the state. It also looked at some components of direct democracy in Switzerland. Therefore, the paper focused on direct democracy, using secondary sources of data. In conclusion, the research offers direct democracy as a solution to the failure of the Nigerian administrative system especially as it affects participation, developmental programmes and institutionalized corruption.

Keywords: corruption, direct democracy, national development, Nigeria, participation

Procedia PDF Downloads 446

Authors: Prachi Choudhary, Jai Gopal Sharma

Abstract:

Microbial cultures determine the type of organism, its abundance in the tested sample, or both. It is one of the primary diagnostic methods of microbiology. It is used to determine the cause of infectious disease by letting the agent multiply in a predetermined medium. Different bacteria produce colonies that may be very distinct from the bacterial species that produced them. To culture any pathogen or microorganism, we should first know about the types of media used in microbiology for culturing. Sometimes sub culturing is also done in various microorganisms if some mixed growth is seen in culture. Nearly 3 types of culture media based on consistency – solid, semi-solid, and liquid (broth) media; are further explained in the report. Then, The Five I's approach is a method for locating, growing, observing, and characterizing microorganisms, including inoculation and incubation. Isolation, inspection, and identification. For identification of bacteria, we have to culture the sample like urine, sputum, blood, etc., on suitable media; there are different methods of culturing the bacteria or microbe like pour plate method, streak plate method, swabbing by needle, pipetting, inoculation by loop, spreading by spreader, etc. After this, we see the bacterial growth after incubation of 24 hours, then according to the growth of bacteria antibiotics susceptibility test is conducted; this is done for sensitive antibiotics or resistance to that bacteria, and also for knowing the name of bacteria. Various methods like the dilution method, disk diffusion method, E test, etc., do antibiotics susceptibility tests. After that, various medicines are provided to the patients according to antibiotic sensitivity and resistance.

Keywords: inoculation, incubation, isolation, antibiotics suspectibility test, characterizing

Procedia PDF Downloads 45

Authors: Muhammad Naeem Chaudhry, Fahim Nawaz, Rana Nauman Shabbir

Abstract:

New strategies aimed at increasing the resilience of crop plants to the negative effects of climate change represent important research priorities of plant scientists. The use of soil microorganisms to alleviate abiotic stresses like drought has gained particular importance in recent past. A field experiment was planned to investigate the effect of phosphorous solubilizing bacteria on yield and seed quality of Camelina (Camelina sativa L.) under water deficit conditions. The study was conducted at Agronomic Research Farm, University College of Agriculture and Environmental Sciences, The Islamia University Bahawalpur, during 4th week of November, 2013. The available seeds of Camelina sativa were inoculated with two bacterial strains (pseudomonas and Bacillus spp.) and grown under various water stress levels i.e. D0, (four irrigations), D3 (three irrigation), D2 (two irrigations), and D1 (one irrigation). The results revealed that drought stress significantly reduced the plant growth and yield, consequently reducing protein contents and oil concentration in camelina. The exposure to drought stress decreased plant height (16%), plant population (27%), number of fertile branches (41-59%), number of pods per plant (35%) and seed per pod (33%). Drought stress also exerted a negative impact on yield characteristics by reducing the 1000-seed weight (65%), final seed yield (52%), biological yield (22%) and harvest index (39%) of camelina. However, the inoculation of seeds with Pseudomonas and Bacillus spp. promoted the plant growth characterized by increased plant height and enhanced plant population. It was noted that inoculation of seeds with Pseudomonas resulted in the maximum plant population (113.4 cm), primary branches (19 plant-1), and number of pods (664 plant-1), whereas Bacillus inoculation resulted in maximum plant height (113.4 cm), seeds per pod (15.9), 1000-seed weight (1.85 g), and seed yield (3378.8 kg ha-1). Moreover, the inoculation with Bacillus also significantly improved the quality attributes of camelina and gave 3.5% and 2.1% higher oil contents than Pseudomonas and control (no-inoculation), respectively. Similarly, the same strain also resulted in maximum protein contents (33.3%). Our results confirmed the hypothesis that inoculation of seeds with phosphorous solubilizing bacterial strains is an effective, viable and environment-friendly approach to improve yield and quality of camelina under water deficit conditions. However, further studies are suggested to investigate the physiological and molecular processes, stimulated by bacterial strains, for increasing drought tolerance in food crops.

Keywords: Camelina, drought stress, phosphate solubilizing bacteria, seed quality

Procedia PDF Downloads 235

Authors: H. Kishor, G. Prabhuling, D. S. Ambika, D. P. Prakash

Abstract:

The chemical mutagens have become important tool to enhance agronomic traits of banana crop. It is being used to develop fusarium resistance lines in various susceptible banana cultivars. There are several mutagens like EMS and NaN3 available for banana crop improvement and each mutagen has its own important role as positive or negative effects on growth and development of banana plants. Explants from shoot tip culture were treated with various EMS (0.30, 0.60, 0.90 and 0.12%) and NaN3 (0.01, 0.02 and 0.03%) concentrations. The putative mutants obtained after in vitro rooting were subjected for artificial inoculation of Fusarium oxysporum f.sp. cubense. Screening putative mutants resistance to Panama disease was carried out by using syringe method of inoculation. It was observed that, EMS treated mutants were more susceptible compared to NaN3 treatment. Among the NaN3 doses 0.01% found to produce 3 resistant lines during preliminary screening under greenhouse conditions.

Keywords: Nanjangud rasabale, EMS, NaN3, putative mutants

Procedia PDF Downloads 156

Authors: M. Ottenbrite, G. Yilmaz, J. Devenish, M. Kang, H. Dan, M. Lin, C. Lau, C. Carrillo, K. Bessonov, J. Nash, E. Topp, J. Guan

Abstract:

Consumption of food contaminated by antibiotic-resistant (AR) bacteria may lead to the transmission of AR genes in the gut microbiota and cause AR bacterial infection, a significant public health concern. However, information is limited on if and how background microbes from the food matrix (food microbes) may influence resistance transmission. Thus, we assessed the colonization of a β-lactam resistant Salmonella Heidelberg strain (donor) and a β-lactam susceptible S. Typhimurium strain (recipient) and the transfer of the resistance genes in the mouse gut in the presence or absence of food microbes that were derived from washing freshly-harvested carrots. Mice were pre-treated with streptomycin and then inoculated with both donor and recipient bacteria or recipient only. Fecal shedding of the donor, recipient, and transconjugant bacteria was enumerated using selective culture techniques. Transfer of AR genes was confirmed by whole genome sequencing. Gut microbial composition was determined by 16s rRNA amplicon sequencing. Significantly lower numbers of donors and recipients were shed from mice that were inoculated with food microbes compared to those without food microbe inoculation. S. Typhimurium transconjugants were only recovered from mice without inoculation of food microbes. A significantly higher survival rate was in mice with vs. without inoculation of food microbes. The results suggest that the food microbes may compete with both the donor and recipient Salmonella, limit their growth and reduce transmission of the β-lactam resistance gene in the mouse gut.

Keywords: antibiotic resistance, gene transfer, gut microbiota, Salmonella infection

Procedia PDF Downloads 38

Authors: Alise Senberga, Laila Dubova, Liene Strauta, Ina Alsina, Ieva Erdberga

Abstract:

Legume symbiotic relationship with nitrogen fixating bacteria Rhizobim leguminosarum is an important factor used to improve the productivity of legumes, due to the fact that rhizobia can supply plant with the necessary amount of nitrogen. R. leguminosarum strains have shown different activity in fixing nitrogen. Depending on the chosen R. leguminosarum strain, host plant biochemical content can be altered. In this study we focused particularly on the changes in protein content in beans (using two different varieties) and peas (five different varieties) due to the use of several different R. leguminosarum strains (four strains for both beans and peas). Overall, the protein content increase was observed after seed inoculation with R. leguminosarum. Strain and plant cultivar interaction specification was observed. The effect of R. leguminosarum inoculation on the content of protein was dependent on the R. leguminosarum strain used. Plant cultivar also appeared to have a decisive role in protein content formation with the help of R. leguminosaru.

Keywords: legumes, protein content, rhizobia strains, soil

Procedia PDF Downloads 482

Authors: R. Alias, A. Kasa, M. R. Taha

Abstract:

The effect of particle size on shear strength of granular materials are investigated using direct shear tests. Small direct shear test (60 mm by 60 mm by 24 mm deep) were conducted for particles passing the sieves with opening size of 2.36 mm. Meanwhile, particles passing the standard 20 mm sieves were tested using large direct shear test (300 mm by 300 mm by 200 mm deep). The large direct shear tests and the small direct shear tests carried out using the same shearing rate of 0.09 mm/min and similar normal stresses of 100, 200, and 300 kPa. The results show that the peak and residual shear strength decreases as particle size increases.

Keywords: particle size, shear strength, granular material, direct shear test

Procedia PDF Downloads 454

Authors: Saad Bin Nasir

Abstract:

The study identifies the impact of inflation and foreign exchange rate on foreign direct investment in Pakistan. Inflation and exchange rates are used as independent variables and foreign direct investment is taken as dependent variable. Discreet time series data has been used from the period of 1999 to 2009. The results of regression analysis reveal that high inflation has negative impact on foreign direct investment and higher exchange rates has positive impact on foreign direct investment in Pakistan. The inflation and foreign exchange rates both are insignificant in the analysis.

Keywords: inflation rate, foreign exchange rate, foreign direct investment, foreign assets

Procedia PDF Downloads 383

Authors: Hanane Boutaj, Abdelilah Meddich, Said Wahbi, Zainab El Alaoui-Talibi, Allal Douira, Abdelkarim Filali-Maltouf, Cherkaoui El Modafar

Abstract:

The present work aims to investigate the effect of arbuscular mycorrhizal fungi (AMF) in improving the olive tree resistance to Verticillium wilt caused by Verticillium dahliae. Inoculated plants with a mycorrhizal autochthonous consortium 'Rhizolive consortium' and pure strain 'Glomus irregulare' were infected after three months with V. dahliae. The improving of olive tree resistance was determined through disease severity, incidence, and defoliation. On the other hand, the defense mechanisms of olive plants were evaluated through lignin content, phenylalanine ammonia lyase (PAL) activity, and polyphenol content. The results revealed that both AMF significantly (p < 0.05) reduced disease development and the rate of defoliation in infected olive plants. Moreover, the contents of lignin were boosted after mycorrhizal inoculation in both the roots and the stems of olive plants, which remained significantly (p < 0.001) higher after the 90th days of V. dahliae inoculation. PAL activity was increased after V. dahliae inoculation in the stems of 'Rhizolive consortium' treatment that were 17 times higher than those in the roots of olive plants. The polyphenol content in the stems was about twice higher than those in the roots. The reduction of disease severity was accompanied by increased levels of lignin content, PAL activity, and polyphenol content, particularly in the stems of olive plants, indicating the strengthening of the olive plant immune system against V. dahliae.

Keywords: olive tree, Mycorrhizal autochthonous consortium, Glomus irregulare, Verticillium dahliae, defense mechanisms

Procedia PDF Downloads 79

Authors: Jhovana Silvia Escobar Ortega, Ines Eugenia Garcia De Salamone

Abstract:

Cover crops are an agri-technological alternative to improve all properties of soils. Cover crops such as oats and rye could be used to reduce erosion and favor system sustainability when they are grown in the same agricultural cycle of the soybean crop. This crop is very profitable but its low contribution of easily decomposable residues, due to its low C/N ratio, leaves the soil exposed to erosive action and raises the need to reduce its monoculture. Furthermore, inoculation with the plant growth promoting rhizobacteria contributes to the implementation, development and production of several cereal crops. However, there is little information on its effects on forage crops which are often used as cover crops to improve soil quality. In order to evaluate the effect of combined inoculation with Azospirillum brasilense and Pseudomonas fluorescens on rhizosphere microbial communities, field experiments were conducted in the west of Buenos Aires province, Argentina, with a split-split plot randomized complete block factorial design with three replicates. The factors were: type of cover crop, inoculation and fertilization. In the main plot two levels of fertilization 0 and 7 40-0-5 (NPKS) were established at sowing. Rye (Secale cereale cultivar Quehué) and oats (Avena sativa var Aurora.) were sown in the subplots. In the sub-subplots two inoculation treatments are applied without and with application of a combined inoculant with A. brasilense and P. fluorescens. Due to the growth of cover crops has to be stopped usually with the herbicide glyphosate, rhizosphere soil of 0-20 and 20-40 cm layers was sampled at three sampling times which were: before glyphosate application (BG), a month after glyphosate application (AG) and at soybean harvest (SH). Community level of physiological profiles (CLPP) and Shannon index of microbial diversity (H) were obtained by multivariate analysis of Principal Components. Also, the most probable number (MPN) of nitrifiers and cellulolytics were determined using selective liquid media for each functional group. The CLPP of rhizosphere microbial communities showed significant differences between sampling times. There was not interaction between sampling times and both, types of cover crops and inoculation. Rhizosphere microbial communities of samples obtained BG had different CLPP with respect to the samples obtained in the sampling times AG and SH. Fertilizer and depth of sampling also caused changes in the CLPP. The H diversity index of rhizosphere microbial communities of rye in the sampling time BG were higher than those associated with oats. The MPN of both microbial functional types was lower in the deeper layer since these microorganisms are mostly aerobic. The MPN of nitrifiers decreased in rhizosphere of both cover crops only AG. At the sampling time BG, the NMP of both microbial types were larger than those obtained for AG and SH. This may mean that the glyphosate application could cause fairly permanent changes in these microbial communities which can be considered bio-indicators of soil quality. Inoculation and fertilizer inputs could be included to improve management of these cover crops because they can have a significant positive effect on the sustainability of the agro-ecosystem.

Keywords: community level of physiological profiles, microbial diversity, plant growth promoting rhizobacteria, rhizosphere microbial communities, soil quality, system sustainability

Procedia PDF Downloads 369

Authors: Joel Ballesteros, Harriet Jane Caleja, Florian Del Mundo, Cherrie Pascual

Abstract:

Diffraction-based sensing was utilized in the quantification of human ferritin in blood serum to provide an alternative to label-based immunoassays currently used in clinical diagnostics and researches. The diffraction intensity was measured by the diffractive optics technology or dotLab™ system. Two methods were evaluated in this study: direct immunoassay and direct sandwich immunoassay. In the direct immunoassay, human ferritin was captured by human ferritin antibodies immobilized on an avidin-coated sensor while the direct sandwich immunoassay had an additional step for the binding of a detector human ferritin antibody on the analyte complex. Both methods were repeatable with coefficient of variation values below 15%. The direct sandwich immunoassay had a linear response from 10 to 500 ng/mL which is wider than the 100-500 ng/mL of the direct immunoassay. The direct sandwich immunoassay also has a higher calibration sensitivity with value 0.002 Diffractive Intensity (ng mL-1)-1) compared to the 0.004 Diffractive Intensity (ng mL-1)-1 of the direct immunoassay. The limit of detection and limit of quantification values of the direct immunoassay were found to be 29 ng/mL and 98 ng/mL, respectively, while the direct sandwich immunoassay has a limit of detection (LOD) of 2.5 ng/mL and a limit of quantification (LOQ) of 8.2 ng/mL. In terms of accuracy, the direct immunoassay had a percent recovery of 88.8-93.0% in PBS while the direct sandwich immunoassay had 94.1 to 97.2%. Based on the results, the direct sandwich immunoassay is a better diffraction-based immunoassay in terms of accuracy, LOD, LOQ, linear range, and sensitivity. The direct sandwich immunoassay was utilized in the determination of human ferritin in blood serum and the results are validated by Chemiluminescent Magnetic Immunoassay (CMIA). The calculated Pearson correlation coefficient was 0.995 and the p-values of the paired-sample t-test were less than 0.5 which show that the results of the direct sandwich immunoassay was comparable to that of CMIA and could be utilized as an alternative analytical method.

Keywords: biosensor, diffraction, ferritin, immunoassay

Procedia PDF Downloads 322