Search results for: incubation

Authors: P. Singh, R. M. Banik

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Artificial neural network (ANN) was employed to optimize assay parameters viz., time, temperature, pH of reaction mixture, enzyme volume and substrate concentration of L-glutaminase from Bacillus cereus MTCC 1305. ANN model showed high value of coefficient of determination (0.9999), low value of root mean square error (0.6697) and low value of absolute average deviation. A multilayer perceptron neural network trained with an error back-propagation algorithm was incorporated for developing a predictive model and its topology was obtained as 5-3-1 after applying Levenberg Marquardt (LM) training algorithm. The predicted activity of L-glutaminase was obtained as 633.7349 U/l by considering optimum assay parameters, viz., pH of reaction mixture (7.5), reaction time (20 minutes), incubation temperature (35˚C), substrate concentration (40mM), and enzyme volume (0.5ml). The predicted data was verified by running experiment at simulated optimum assay condition and activity was obtained as 634.00 U/l. The application of ANN model for optimization of assay conditions improved the activity of L-glutaminase by 1.499 fold.

Keywords: Bacillus cereus, L-glutaminase, assay parameters, artificial neural network

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Authors: Noor-ul-Ain, Muhammad Tahir Khan, Kashif Khan, Adeela Ajmal, Hamid Mustafa

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The objective of this study was to evaluate the preservative and upgrading potential of urea (70g/kg DM) added to high moisture oat silage at laboratory scale trial and urea was hydrolysed 95%. Microbial activity measured by pH and volatile fatty acids (VFA) and lactate production was reduced (p<0.001) by the urea addition. The pH of oat silage (without treated) was measured 5.7 and increased up to 8.00 on average while; volatile fatty acids (VFA) concentration was decreased. Relative proportions of fermentation acids changed after urea addition, increasing the acetate and butyrate and decreasing the propionate and lactate proportions. The addition of urea to oat silages increased (P<0.001) water soluble and ammonium nitrogen of the forage. These nitrogen fractions represented more than 40% of total nitrogen. After urea addition, total nitrogen content of oat silages increased from 21.0 g/kg DM to 28 g/kg DM. Application of urea at a rate of 70 g/kg DM significantly increased (P<0.001) the in situ degradation of neutral-detergent fibre after 48h of rumen incubation (NDF-situ). The NDF-situ was 200 g/kg NDF higher on oat forages ensiled with urea than on oat forages ensiled without urea. Oat silages can be effectively preserved and upgraded by ensiling with 70 g urea/kg dry matter. Further studies are required to evaluate voluntary intake of this forage.

Keywords: oat, silage, urea, pH, forage

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Authors: Hamdi Belfeki, Belgacem Chandoul, Mnasser Hassouna, Mondher Mejri

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Aqueous and ethanolic extracts of eight Tunisian aromatic and medicinal plants (TAMP) were characterized by studying their composition in polyphenols and also their antiradical and antioxidant capacities. In absence and in the presence of the various extracts, α-amylase from Bacillus subtlis activity, was measured in order to detect a potential inhibition. The total contents of polyphenols and flavonoid vary in function of TAMP and the mobile phase used for the extraction (distilled water or ethanol). The ethanolic extracts showed the most significant antiradical and antioxidant activities. Only the extracts from Coriandrum sativum showed a significant inhibiting effect on the α-amylase activity. This inhibiting capacity could be correlated with the chemical profile of the two extracts, due to the fact that they have the greatest amount of total flavonoid. The ethanolic extract has the most important antioxidant and anti-radicalizing activities among the sixteen extracts studied. The inhibition kinetics of the two coriander extracts were evaluated by pre-incubation method, using Lineweaver-Burk’s equation, obtained by linearization of Michaeilis-Menten’s expression. The results showed that both extracts exercised a competitive inhibition mechanism.

Keywords: α-amylase, antioxidant activity, aromatic and medicinal plants, inhibition

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Authors: Bipul Behari Saha, Sunil Kumar Singh, P. Padmaja, Kamlesh Vishwakarma

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Dissipation study of tebuconazole in alluvial, black and deep-black clayey soils collected from paddy, mango and peanut cropland of tropical agro-climatic zone of India at three concentration levels were carried out for monitoring the water contamination through persisted residual toxicity. The soil-slurry samples were analyzed by capillary GC-NPD methods followed by ultrasound-assisted extraction (UAE) technique and cleanup process. An excellent linear relationship between peak area and concentration obtained in the range 1 to 50 μgkg-1. The detection (S/N, 3 ± 0.5) and quantification (S/N, 7.5 ± 2.5) limits were 3 and 10 μgkg-1 respectively. Well spiked recoveries were achieved from 96.28 to 99.33 % at levels 5 and 20 μgkg-1 and method precision (% RSD) was ≤ 5%. The soils dissipation of tebuconazole was fitted in first order kinetic-model with half-life between 34.48 to 48.13 days. The soil organic-carbon (SOC) content correlated well with the dissipation rate constants (DRC) of the fungicide Tebuconazole. An increase in the SOC content resulted in faster dissipation. The results indicate that the soil organic carbon and tebuconazole concentrations plays dominant role in dissipation processes. The initial concentration illustrated that the degradation rate of tebuconazole in soils was concentration dependent.

Keywords: cropland soil, dissipation, laboratory incubation, tebuconazole

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Authors: Khaled M. Khleifat

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An extracellular methanol and ethanol tolerant lipase producing bacterial strain K5b4 was isolated from soil samples contaminated with hydrocarbon residues. It was identified by using morphological and biochemical characteristics and 16srRNA technique as Acinetobacter species. The immobilized lipase from Acinetobacter sp. K5b4 retained more than 98% of its residual activity after incubation with pure methanol and ethanol for 24 hours. The highest hydrolytic activity of the immobilized enzyme was obtained in the presence of 75% (v/v) methanol in the assay solution. In contrary, the enzyme was able to maintain its original activity up to only 25% (v/v) ethanol whereas at elevated concentrations of 50 and 75% (v/v) the enzyme activity was reduced to 10 and 40%, respectively. Maximum lipase activity of 31.5 mU/mL was achieved after 48 hr cultivation when the optimized medium (pH 7.0) that composed of 1.0% (w/v) olive oil, 0.2% (w/v) glycerol, 0.15% (w/v) yeast extract, and 0.05% (w/v) NaCl was inoculated with 0.4% (v/v) seed culture and incubated at 30°C and 150 rpm agitation speed. However, the presence of CaCl2 in the growth media did not show any inhibitory or stimulatory effect on the enzyme production as it compared to the control experiment. Meanwhile, the other mineral salts MgCl2, MnCl2, KCl and CoCl2 were negatively affected the production of lipase enzyme. The inhibition of lipase production from Acinetobacter sp. K5b4 in presence of glucose suggesting that lipase gene expression is prone to catabolic repression.

Keywords: K5B4, methanol and ethanol, acinetobacter, morphological

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Authors: D. P. Rollinson, B. J. Dilley, P. G. Ryan

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The white-chinned petrel (Procellaria aequinoctialis) is the seabird species most commonly killed by Southern Hemisphere longline fisheries. Despite the importance of diving ability for mitigating longline bycatch, little is known of this species’ diving behaviour. We obtained data from temperature–depth recorders from nine white-chinned petrels breeding on Marion Island, southwestern Indian Ocean, during the late incubation and chickrearing period. Maximum dive depth (16 m) was slightly deeper than the previous estimate (13 m), but varied considerably among individuals (range 2–16 m). Males dived deeper than females, and birds feeding chicks dived deeper than incubating birds, but dive rate did not differ between the sexes. Time of day had no significant effect on dive depth or rate. Our findings will help to improve the design and performance of mitigation measures aimed at reducing seabird bycatch in longline fisheries, such as the calculation of minimum line sink rates and optimum aerial coverage of bird-scaring lines.

Keywords: dive depth, dive duration, temperature–depth recorders, seabirds, bird-scaring lines

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Authors: Surendra Bodakhe

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In this investigation, anticataract activity was determined using cataract formation in developing chick embryo by hydrocortisone. Lenses were evaluated firstly for the extent of opacity and secondly, for lens glutathione (GSH) levels. Betulinic acid was isolated from the chloroform fraction of the crude ethanolic extract of Bauhinia variegata bark (SBE). Fourteen days old Australorp fertilized eggs were divided into different groups of six eggs each. After 24 hrs incubation in a humidified incubator (37οC), at 15 days of age; hydrocortisone (0.25µM/0.2ml/egg) was administered to the chorioallantoic membrane of chick embryos through a small hole in the egg shell on the air sack. Ascorbic acid (standard) or Betulinic acid (test) were administered at 3, 10 and 20 hr after hydrocortisone administration at a specified dose. The puncture was sealed with a cellophane tape and eggs were incubated for 48 hrs in a humidified incubator at 37οC. After 48 hrs, the lenses were isolated for the determination of the extent of opacity and Glutathione level. The betulinic acid prevented the opacification of the chick embryo lenses induced by hydrocortisone. The betulinic acid also prevented the decline of GSH content caused by hydrocortisone. The results indicate that betulinic acid protect the cataract formation in chick embryo lenses induced by hydrocortisone.

Keywords: betulinic acid, cataract, cloudiness, ovine

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Authors: Huyuan Zhang, Yi Chen

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Compacted sewage sludge has been proved to be feasible as a barrier material for tailing impoundment because of its low permeability and retardation of heavy metals. The long-term penetration of acid mine drainage, however, would acidify the barrier system and result in remobilization of previously immobilized heavy metal pollutants. In this study, the effect of decreasing pH on the mobility of three typical heavy metals (Zn, Pb, and Cu) is investigated by acid titration test on sewage sludge under various conditions. The remobilization of heavy metals is discussed based on the acid buffering capacity of sewage sludge-leachate system. Test results indicate that heavy metals are dramatically released out when pH is decreased below 6.2, and their amounts take the order of Zn > Cu > Pb. The acid buffering capacity of sewage sludge decreases with the solid-liquid ratio but increases with the anaerobic incubation time, and it is mainly governed by dissolution of contained carbonate and organics. These results reveal that the sewage sludge possesses enough acid buffering capacity to consume protons within the acid mine drainage. Thus, this study suggests that an explosive remobilization of heavy metals is not expected in a long-term perspective.

Keywords: acid buffering capacity, barrier, heavy metals, remobilization, sewage sludge

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Authors: Yetti Marlida, Syukri Arif, Nadirman Haska

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Recently, alcohol fuels have been produced on industrial scales by fermentation of sugars derived from wheat, corn, sugar beets, sugar cane etc. The enzymatic hydrolysis of cellulosic materials to produce fermentable sugars has an enormous potential in meeting global bioenergy demand through the biorefinery concept, since agri-food processes generate millions of tones of waste each year (Xeros and Christakopoulos 2009) such as sugar cane baggase , wheat straw, rice straw, corn cob, and oil palm trunk. In fact oil palm trunk is one of the most abundant lignocellulosic wastes by-products worldwide especially come from Malaysia, Indonesia and Nigeria and provides an alternative substrate to produce useful chemicals such as bioethanol. Usually, from the ages 3 years to 25 years, is the economical life of oil palm and after that, it is cut for replantation. The size of trunk usually is 15-18 meters in length and 46-60 centimeters in diameter. The trunk after cutting is agricultural waste causing problem in elimination but due to the trunk contains about 42% cellulose, 34.4%hemicellulose, 17.1% lignin and 7.3% other compounds,these agricultural wastes could make value added products (Pumiput, 2006).This research was production of bioethanol from oil palm trunk via saccharafication by cocktail carbohydrases enzymes. Enzymatic saccharification of acid treated oil palm trunk was carried out in reaction mixture containing 40 g treated oil palm trunk in 200 ml 0.1 M citrate buffer pH 4.8 with 500 unit/kg amylase for treatment A: Treatment B: Treatment A + 500 unit/kg cellulose; C: treatment B + 500 unit/kgg xylanase: D: treatment D + 500 unit/kg ligninase and E: OPT without treated + 500 unit/kg amylase + 500 unit/kg cellulose + 500 unit/kg xylanase + 500 unit/kg ligninase. The reaction mixture was incubated on a water bath rotary shaker adjusted to 600C and 75 rpm. The samples were withdraw at intervals 12 and 24, 36, 48,60, and 72 hr. For bioethanol production in biofermentor of 5L the hydrolysis product were inoculated a loop of Saccharomyces cerevisiae and then incubated at 34 0C under static conditions. Samples are withdraw after 12, 24, 36, 48 and 72 hr for bioethanol and residual glucose. The results of the enzymatic hidrolysis (Figure1) showed that the treatment B (OPT hydrolyzed with amylase and cellulase) have optimum condition for glucose production, where was both of enzymes can be degraded OPT perfectly. The same results also reported by Primarini et al., (2012) reported the optimum conditions the hydrolysis of OPT was at concentration of 25% (w /v) with 0.3% (w/v) amylase, 0.6% (w /v) glucoamylase and 4% (w/v) cellulase. In the Figure 2 showed that optimum bioethanol produced at 48 hr after incubation,if time increased the biothanol decreased. According Roukas (1996), a decrease in the concentration of ethanol occur at excess glucose as substrate and product inhibition effects. Substrate concentration is too high reduces the amount of dissolved oxygen, although in very small amounts, oxygen is still needed in the fermentation by Saccaromyces cerevisiae to keep life in high cell concentrations (Nowak 2000, Tao et al. 2005). The results of the research can be conluded that the optimum enzymatic hydrolysis occured when the OPT added with amylase and cellulase and optimum bioethanol produced at 48 hr incubation using Saccharomyses cerevicea whereas 18.08 % bioethanol produced from glucose conversion. This work was funded by Directorate General of Higher Education (DGHE), Ministry of Education and Culture, contract no.245/SP2H/DIT.LimtabMas/II/2013

Keywords: oil palm trunk, enzymatic hydrolysis, saccharification

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Authors: Ghoul Rachid Brahim, Trad Khodja Rafik

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Vertebral localization of the hydatid cyst is a severe form of bone hydatidosis, is a parasitic infection caused by the larval forms of the tapeworms Echinococcus granulosus, The disease is slowly remaining silent (a long incubation period) which may explain why this pathology is often discovered at the stage of neurological complications. The objective of this study is to recall the clinical and radiological aspects of this condition and the importance of early diagnosis and appropriate management. We report a study of 5 patients with vertebral hydatidosis, four men and one woman, four (04) patients operated in the emergency setting for spinal cord compression (decompression by wide laminectomy with evacuation of intra and extra canal vesicles).Albendazole-based medical treatment is instituted in all patients. Results: The evolution was favorable for three patients, the other two patients reoperated for a local recurrence. Conclusion: Vertebral hydatidosis is a rare condition with a poor prognosis due to the risk of neurological damage, the infiltrating nature of bone lesions, the frequency of relapses and therapeutic difficulties. The only curative method remains surgery, which must aim for complete and large excision of the lesions as if it were a “malignant tumour”.

Keywords: hydatidosis, Echinococcosis granulosus, hydatid cyst, spinal cord compression, laminectomy

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Authors: Isaac Samuel

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A study was conducted to determine the in-vitro degradation of selected Nigerian browse plants consumed by small ruminants on free range in northern guinea savannah region of Nigeria using in vitro gas production, proximate composition, fibre components, methane gas production and dry matter degradation as tools. The leaves samples of the selected browse plants were collected, processed and incubated using in vitro gas dry matter degradation techniques. Results obtained showed variation in the rate of degradation. The result obtained from chemical analysis showed that the CP content of A. occidentale (26.49%) was higher than F. thonningi (23.58%), M. indica (20.58%) and T. catappa (18.61%). Both ADF and NDF of A. occidentale (40.00 and 50.00) were as well higher than F. thonningi (20.00 and 40.00), M. indica (20.00 and 40.00) and T.catappa (20.00 and 42.00). Results from in vitro gas production however showed that T. catappa (23.67ml/DM) has a significantly higher (p<0.05) value than F.thonningi (20.67ml/DM), A. occidentale (16.67ml/DM), and M. indica(14.00ml/DM) at 72 hours of incubation. Methane gas production and in vitro gas production can be used to predict dry matter degradation and nutritive value of feedstuff for small ruminants. A. occidentale with the least methane gas production and highest crude protein (CP) content might have the most nutritive value among the browse plants investigated.

Keywords: in vitro, degradation, browse, gas production

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Authors: Rasha Saad, Jean Frederic Weber, Fatimah Bebe, Sadia Sultan

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The goal of study was to screen the effects of candesartan and four endophytic fungi for their potential in microbial biotransformation. In this experiment, four types of unidentified fungi with the codes of TH2L1, TH2R10, TH1P35 and TH1S46 were used in screening process by MECFUS (Microtiter plate, Elicitors, Combination, Freeze-drying, UHPLC, Statistical analysis) protocol. The experiment was carried out by using 96-well microtiter plate (MTP) with different media and elicitors. Various media with two concentrations of Potato Dextrose Broth (PDB) and elicitors used were to induce the production of secondary metabolites from the fungi as well as the biotransformation of the drug compound. After incubation, cultures were extracted by freeze drying method and finally analyzed by ultra-High performance Liquid Chromatography (uHPLC). The extracts analyzed by uHPLC followed by LC/Ms, demonstrated the presence of biotransformation products from the drug compound and elicitation of the secondary metabolism from the fungi by the occurrence of the additional peaks. From the four fungi, TH1S46 showed highly potential produced secondary metabolites as well as the biotransformation of candesartan. For other fungi, they responded when candesartan was introduced. Moreover, the additional peaks produced in uHPLC need to be further investigation by using LC-MS or NMR.

Keywords: biotransformation, candesartan, endophytes, secondary metabolites

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Authors: Hadriana Bansi, Elizabeth Wina, Toto Toharmat

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Acacia villosa is thornless shrub legume which contents high crude protein. However, the utilization of A. villosa as ruminant feed is limited by its secondary compounds. The aim of this article is to find out the maximum of substitution A. villosa in sheep ration. The nutritional evaluation consisted of in vitro two stages, in vivo, and in vitro gas production trials. The secondary compounds of A. villosa also were analyzed. Evaluating digestibility of increasing level of substitution A. villosa replacing Pennisetum purpureum was using in vitro two stages. The substitution of 30% A. villosa was compared to 100% P. purpureum by in vitro gas production technique and in vivo digestibility. The results of two stages in vitro showed that total phenol, condensed tannin, and non-protein amino acid (NPAA) were high. Substitution 15% A. villosa reached the highest digestibility for both dry matter (DM) and crude protein (CP) which were 67% and 86% respectively, but it was shown that DM and CP digestibility of substitution 30% of A. villosa was still high which were 61.82% and 75-67% respectively. The pattern of gas production showed that first 8 hours total gas production substitution of 30% A. villosa was higher than 100% P. purpureum and declined after 10 hours incubation. In vivo trials showed that substitution of 30% A. villosa significantly increased CP intake, CP digestibility, and nitrogen retention. It can be concluded that substitution A. villosa until 30% still gave the good impact even though it has high secondary compounds.

Keywords: Acacia villosa, digestibility, gas production, secondary compounds

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Authors: Stephen Adeniyi Adefegha, Vitor Mostardeiro, Vera Maria Morsch, Ademir F. Morel, Ivana Beatrice Manica Da Cruz, Sabrina Somacal Maria Rosa Chitolina Schetinger

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Moringa stenopetala is often consumed as food and used in folkloric medicine for the management of several diseases. Purpose: This study was set up in order to assess the effect of aqueous extract of Moringa stenopetala on cell viability and oxidative stress biomarkers in BV-2 microglial cells. Aqueous extracts of M. stenopetala were prepared, lyophilized and reconstituted in 0.5% dimethylsulphoxide (DMSO). Cells were treated with M. stenopetala extracts (0.1 - 100 µg/ml) for cell viability and nitric oxide (NO) production tests. However, M. stenopetala extract (50 µg/ml) was used in the treatment of cells for the determination of protein carbonyl content and reactive oxygen species (ROS) level. Incubation of BV-2 microglia cell with M. stenopetala extract maintained cell viability, diminished NO and ROS levels, and reduced protein carbonyl contents Chlorogenic acid, rutin, kaempferol and quercetin derivatives were the main phenolic compounds identified in M. stenopetala leaf extract. These phenolic compounds present in M. stenopetala may be responsible for the mitigation of oxidative stress in BV-2 microglial cells.

Keywords: oxidative stress, BV-2 microglial cell, Moringa stenopetala, cell viability, antioxidant

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Authors: Fatemeh Aghamohammadzadeh, Mahdi Asghari Ozma

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SARS-CoV-2 was transmitted from animals to humans in China and through air transport to almost all world countries, including Iran, creating the first pandemic of the 21st century. The virus was spread through droplets from sneezing, coughing, loud talking, and exhalation of sick and asymptomatic people, even during incubation. It was transmitted from human to human directly by inhalation of viruses in droplets or indirectly through contact with infected surfaces, resulting in the death of a significant number of patients, especially the elderly and those with underlying diseases. The virus is more likely to be transmitted in places with high population densities. The chain of transmission of infection can be broken by observing the following: risk perception, reduced travel, complete quarantine in a particular area, home quarantine, social distancing, use of personal protective equipment (PPE), prevention of gatherings, cleaning and disinfection of public utilities and busy places, identifying, isolating and treating infected people, tracking calls, continuing health education, following health principles by people, especially in poor areas, and washing their hands frequently with soap and water or disinfecting them with 70% ethanol.

Keywords: COVID-19, transmission, population density, home quarantine, social distancing

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Authors: Maged Syed Ahamd, Manal Mohamed Yasser, Essam Sholkamy

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In this paper, we reported that selenium (Se) nanoparticles were firstly biosynthesized with a simple and eco-friendly biological method. Their shape, size, FTIR (Fourier Transform Infrared spectroscopy), UV–vis spectra, TEM (Transmission Electron Microscopy) images and EDS (Energy Dispersive Spectroscopy) pattern have been analyzed. TEM analyses of the samples obtained at different stages indicated that the formation of these Se nanostructures was governed by an incubation time (12- 24- 48 hours). The Se nanoparticles were initially generated and then would transform into crystal seeds for the subsequent growth of nanowires; however obtaining stable Se nanowire with a diameter of about 15-100 nm. EDS shows that Se nanoparticles are entirely pure. The IR spectra showed the peaks at 550 cm-1, 1635 cm-1, 1994 cm-1 and 3430 cm-1 correspond to the presence of Se-O bending and stretching vibrations. The concentrations of Se-NPs (0, 1, 2, 5 µg/ml) did not give significantly effect on both two cell lines while the highest concentrations (10- 100 µg/ml gave significantly effects on them. The lethal dose (ID50%) of Se-NPs on Hep2 G and MCF-7 cells was obtained at 75.96 and 61.86 µg/ml, respectively. Results showed that Se nanoparticles as anticancer agent against MCF-7 cells were more effective than Hep2 G cells. Our results suggest that Se-NPs may be a candidate for further evaluation as a chemotherapeutic agent for breast and liver cancers.

Keywords: selenium nanoparticle, Streptomyces bikiniensis, nanowires, chemotherapeutic agent

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Authors: Sepehr Moradi, Mehdi Asadi Rad

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The aim of this study is to evaluate and compare chest weight of pure and mixed races Kabood 30-day Pigeons to investigate about their sex, race, and some auxiliary variables. In this paper, 62 pieces of pigeons as 31 male and female pairs with equal age are studied randomly. A natural incubation was done from each pair. All produced chickens were slaughtered at 30 days age after 12 hours hunger. Then their chests were weighted by a scale with one gram precision. A covariance analysis was used since there were many auxiliary variables and unequal observations. SAS software was used for statistical analysis. Mean weight of chests in pure race (Kabood-Kabood) with 8 records, 123.8±32.3g and mixed races of Kabood-Namebar, Kabood-Parvazy, Kabood-Tizpar, Namebar-Kabood, Tizpar-Kabood, and Parvazi-Kabood with 8, 8, 6, 12, 10, and 10 records were 139.4±23.5, 7/122±23.8, 124.7±30.1, 50.3±29.3, 51.4±26.4, and 137±28.6 gr, respectively. Mean weight of 30-day chests in male and female sex were 87.3±2.5 and 82.7±2.6g, respectively. Difference chest weight of 30-day chests of Kabood-Kabood race with Kabood-Namebar, Kabood-Parvazi, Tizpar-Kabood, Kabood-Tizpar, Namebar-Kabood and Parvazi-Kabood mixed races was not significant. Effect of sex was also significant in 5% level (P<0.05), but mutual effect of sex and race was not significant. Auxiliary variable of father weight was significant in 1% level (p < 0.01), but auxiliary variable of mother weight was not significant. The results showed that most and least weights belonged to Kabood-Namebar and Namebar-Kabood.

Keywords: squab, Kabood race, 30-day chest weight, pigeons

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Authors: Funmilola O. Omoya

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Muscid flies are known to be vectors of disease agents and species that annoy humans and domesticated animals. An example of these flies is Musca domestica (house fly) whose adult and immature stages occur in a variety of filthy organic substances including household garbage and animal manures. They contribute to microbial contamination of foods. It is therefore imperative to control these flies as a result of their role in Public health. The second and third instars of Musca domestica (Linn) were infected with varying cell loads of Bacillus subtilis in vitro for a period of 48 hours to evaluate its larvicidal activities. Mortality of the larvae increased with incubation period after treatment with the varying cell loads. Investigation revealed that the second instars larvae were more susceptible to treatment than the third instars treatments. Values obtained from the third instar group were significantly different (P0.05) from those obtained from the second instars group in all the treatments. Lethal concentration (LC50) at 24 hours for 2nd instars was 2.35 while LC50 at 48 hours was 4.31.This study revealed that Bacillus subtilis possess good larvicidal potential for use in the control of Musca domestica in poultry farms.

Keywords: Bacillus subtilis, Musca domestica, larvicidal activities, poultry farms

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Authors: N. Cetinkaya, Y. S. Kuleyin

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The aim of this study was to estimate the digestibility of the fruit internal skin of different varieties of hazelnuts to propose hazelnut fruit skin as an alternative feed source as roughage in ruminant nutrition. In 2015, the fruit internal skins of three different varieties of round hazelnuts (RH), pointed hazelnuts (PH) and almond hazelnuts (AH) were obtained from hazelnut processing factory then their crude nutrients analysis were carried out. Organic matter digestibility (OMD) and metabolisable energy (ME) values of hazelnut fruit skins were estimated from gas measured by in vitro gas production method. Their antioxidant activities were determined by spectrophotometric method. Crude nutrient values of three different varieties were; organic matter (OM): 87.83, 87.81 and 87.78%), crude protein (CP): 5.97, 5.93 and 5.89%, neutral detergent fiber (NDF): 30.30, 30.29 and 30.29%, acid detergent fiber (ADF): 48.68, 48.67 and 48.66% and acid detergent lignin (ADL): 25.43, 25.43 and 25.39% respectively. OMD from 24 h incubation time of RH, PH and AH were 22.04, 22.46 and 22.74%; ME_GP values were 3.69, 3.75 and 3.79 MJ/kg DM; and antioxidant activity values were 94.60, 94.54 and 94.52 IC 50 mg/mL respectively. The fruit internal skin of different varieties of hazelnuts may be considered as an alternative roughage for ruminant nutrition regarding to their crude and digestible nutritive values. Moreover, hazelnut fruit skin has a rich antioxidant content so it may be used as a feed additive for both ruminant and non-ruminant animals.

Keywords: antioxidant activity, hazelnut fruit skin, metabolizable energy, organic matter digestibility

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Authors: Sankar Majumder

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Development of Rural Entrepreneurs requires a holistic approach involving social, economic, political, technical, and environmental and many other issues. It needs a thorough understanding of the economy and society. It's true that agricultural development, rural development and many other social and right based development programmes have resulted in the growth of income in the rural sector. The development of rural entrepreneurs is necessary to utilise these opportunities. Many programmes and policies in the spheres of organisational, financial, infrastructural and technical supports have been taken to promote rural industries. But if one looks at the growth and development of rural industrial units, especially the manufacturing units, the picture is not promising. This paper aims at analysing the possible causes and its solutions in terms of (1) Mind set of the society towards business as a livelihood; (2) Sufficiency and appropriateness of the existing organisational, financial, infrastructural and technical supports. The paper is based on secondary data on various aspects of rural enterprises and the author’s experiences in the course of his work as a practitioner in this field. Growth of units and employment in the rural industries shows that the entrepreneurs are more inclined towards trading units than towards manufacturing ventures. The growth of rural industries is constrained not by the insufficiency of the supply of finance but by the insufficient demand for finance. The task is to increase the supply of entrepreneurs by creating an entrepreneurial environment. Incubation for rural entrepreneurs is the need of the hour.

Keywords: business mind set, entrepreneurial environment, supply of finance, technical support

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Authors: Ayoub Azizi-Shotorkhoft, Tahereh Mohammadabadi, Hosein Motamedi, Morteza Chaji, Hasan Fazaeli

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This study was conducted to investigate nutritive value of wheat straw processed with termite gut symbiotic bacteria with lignocellulosic-degrading potential including Bacillus licheniformis, Ochrobactrum intermedium and Microbacterium paludicola in vitro. These bacteria were isolated by culturing termite guts contents in different culture media containing different lignin and lignocellulosic materials that had been prepared from water-extracted sawdust and wheat straw. Results showed that incubating wheat straw with all of three isolated bacteria increased (P<0.05) acid-precipitable polymeric lignin (APPL) compared to control, and highest amount of APPL observed following treatment with B. licheniformis. Highest and lowest (P<0.05) in vitro gas production and ruminal organic matter digestibility were obtained when treating wheat straw with B. licheniformis and control, respectively. However, other fermentation parameters such as b (i.e., gas production from the insoluble fermentable fractions at 144h), c (i.e., rate of gas production during incubation), ruminal dry matter digestibility, metabolizable energy, partitioning factor, pH and ammonia nitrogen concentration were similar between experimental treatments (P>0.05). It is concluded that processing wheat straw with isolated bacteria improved its nutritive value as ruminants feed.

Keywords: termite gut bacteria, wheat straw, nutritive value, ruminant

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Authors: P. Christodoulou, M. Gerasimou, S. Mantzoukis, N. Varsamis, G. Kolliopoulou, N. Zotos

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Abstract— Purpose: The Intensive Care Unit (ICU) environment is conducive to the growth of microorganisms. A variety of microorganisms gain access to the intravascular area and are transported throughout the circulatory system. Therefore, examination of the catheters used in ICU patients is of paramount importance. Material and Method: The culture medium is a catheter tip, which is enriched with Tryptic soy broth (TSB). After one day of incubation, the broth is passaged in the following selective media: Blood, Mac conkey No. 2, chocolate, Mueller Hinton, Chapman, and Saboureaud agar. The above selective media is incubated for 2 days. After this period, if any number of microbial colonies is detected, gram staining is performed and then the microorganisms are identified by biochemical techniques in the automated Microscan (Siemens) system followed by a sensitivity test in the same system using the minimum inhibitory concentration (MIC) technique. The sensitivity test is verified by a Kirby Bauer test. Results: In 2017, the Microbiology Laboratory received 84 catheters from the ICU. 42 were found positive. Of these, S. epidermidis was identified at 8, A. baumannii in 10, K. pneumoniae in 6, P. aeruginosa in 6, P. mirabilis in 3, S. simulans in 1, S. haemolyticus in 4, S. aureus in 3 and S. hominis in 1. Conclusions: The results show that the placement and maintenance of the catheters in ICU patients are relatively successful, despite the unfavorable environment of the unit.

Keywords: culture, intensive care unit, microorganisms, vascular catheters

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Authors: Aisami A., Z. A. Adamu, Lawan Bulama

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Due to growing issues of crude oil pollution in both marine and terrestrial environments, Billions to Trillions of US Dollars were spent over the years for the treatment of this spill. There is an urgent need for effective bioremediation strategies. This current study focuses on the isolation and characterization of a crude oil-degrading bacterium from hydrocarbon-contaminated soil samples. Soil samples were collected from an oil spill site and subjected to enrichment culture techniques in a mineral salt medium supplemented with crude oil as the singular carbon source. The isolates were screened for their crude oil-degrading capabilities using gravimetric analysis. The most efficient isolation was identified through 16S rRNA gene sequencing. Cultural and physical conditions such pH, temperature salinity and crude oil concentrations were optimized. The isolates showed significant crude oil degradation efficiency, reducing oil concentration (2.5%) by 75% within 15 days of incubation. The strain was identified as Providencia sp. through molecular characterization, the sequence was deposited at the NCBI Genbank with accession number MN880494. The bacterium exhibited optimal growth at 32.5°C, pH 7.0 to 7.5, and in the presence of 1.5% (w/v) NaCl. The isolated Providencia sp. shows encouraging potential for bioremediation of crude oil-contaminated environments. This study successfully isolated and characterized a crude oil-degrading Providencia sp., highlighting its potential in bioremediation.

Keywords: crude oil degradation, providencia sp., bioremediation, hydrocarbon utilization, environmental pollution.

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Authors: Oktira Roka Aji, Maelita R. Moeis, Ihsanawati, Ernawati A. Giri-Rachman

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Globally, tuberculosis (TB) remains a leading cause of death. The emergence of multidrug-resistant strains and extensively drug-resistant strains have become a major public concern. One of the potential candidates for drug target is the cytoplasmic domain of PhoR Histidine Kinase, a part of the Two Component System (TCS) PhoR-PhoP in Mycobacterium tuberculosis (Mtb). TCS PhoR-PhoP relay extracellular signal to control the expression of 114 virulent associated genes in Mtb. The 3D structure of PhoR cytoplasmic domain is needed to screen novel drugs using structure based drug discovery. The PhoR cytoplasmic domain from Mtb H37Rv was overexpressed in E. coli BL21(DE3), then purified using IMAC Ni-NTA Agarose his-tag affinity column and DEAE-ion exchange column chromatography. The molecular weight of the purified protein was estimated to be 37 kDa after SDS-PAGE analysis. This sample was used for pre-crystallization screening by applying sitting drop vapor diffusion method using Natrix (HR2-116) 48 solutions crystal screen kit at 25ºC. Needle-like crystals were observed after the seventh day of incubation in test solution No.47 (0.1 M KCl, 0.01 M MgCl2.6H2O, 0.05 M Tris-Cl pH 8.5, 30% v/v PEG 4000). Further testing is required for confirming the crystal.

Keywords: tuberculosis, two component system, histidine kinase, needle-like crystals

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Authors: I. R. Keke, C. S. Nwigwe, O. S. Nwanjo, A. S. Egeruoh

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An experiment was carried out at Urban Farm and Fisheries Nigeria Ltd, Owerri Imo State South East Nigeria between February and June 2014 to induce Brood stock of Heterobranchus longifilis (mean wt 1.3kg) in concrete tanks (1.0 x 2.0 x 1.5m) in dimension using a synthetic hormone (Ovaprim) and pituitary extract from Heterobranchus longifilis. Brood stock males were selected as pituitary donors and their weights matched those of females to be injected at 1ml/kg body weight of Fish. Ovaprim, was injected at 0.5ml/kg body weight of female fish. A latency period of 12 hours was allowed after injection of the Brood stock females before stripping the egg and incubation at 23 °C. While incubating the eggs, samples were drawn and the rate of fertilization was determined. Hatching occurred within 33 hours and hatchability rate (%) was determined by counting the active hatchings. The result showed that Ovaprim injected Brood stock eggs fertilized up to 80% while the pituitary from the Heterobranchus longifilis had low fertilization and hatching success 20%. Ovaprim is imported and costly, so more effort is required to enhance the procedures for homoplastic hypophysation.

Keywords: heterobranchus longifilis, ovaprim, hypophysation, latency period, pituitary

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Authors: Osama M. Darwesh, Sahar H. Hassan, Abd El-Raheem R. El-Shanshoury, Shawky Z. Sabae

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Nanotechnology as multidisciplinary technology is growing rapidly with important applications in several sectors. Also, nanobiotechnology is known for the use of microorganisms for the synthesis of targeted nanoparticles. The present study deals with the green synthesis of silver nanoparticles using aquatic bacteria and the development of a biogenic nanocomposite for environmental applications. Twenty morphologically different colonies were isolated from the collected water samples from eight different locations at the Rosetta branch of the Nile Delta, Egypt. The obtained results illustrated that the most effective bacterial isolate (produced the higher amount of AgNPs after 24 h of incubation time) is isolate R3. Bacillus tequilensis was the strongest extracellular bio-manufactory of AgNPs. Biosynthesized nanoparticles had a spherical shape with a mean diameter of 2.74 to 28.4 nm. The antimicrobial activity of silver nanoparticles against many pathogenic microbes indicated that the produced AgNPs had high activity against all tested multi-antibiotic resistant pathogens. Also, the stabilized prepared AgNPs-SA nanocomposite has greater catalytic activity for the decolourization of some dyes like Methylene blue (MB) and Crystal violet. Such results represent a promising stage for producing eco-friendly, cost-effective, and easy-to-handle devices for the bioremediation of contaminated industrial wastewater.

Keywords: bioremediation, AgNPs, AgNPs-SA nanocomposite, Bacillus tequilensis, nanobiotechnology

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Authors: Nadeem A. Siddique, Mohd Mujeeb, Kahkashan

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Introduction: The objective of the projected work is to study the occurrence of the aflatoxins B1, B2, G1and G2 in remedial plants, exclusively in Delphinium denudatum. The aflatoxins were analysed by high-performance liquid chromatography–tandem quadrupole mass spectrometry with electrospray ionization (HPLC–MS/MS) and immunoaffinity column chromatography were used for extraction and purification of aflatoxins. PDA media was selected for fungal count. Results: A good quality linear relationship was originated for AFB1, AFB2, AFG1 and AFG2 at 1–10 ppb (r > 0.9995). The analyte precision at three different spiking levels was 88.7–109.1 %, by means of low per cent relative standard deviations in each case. Within 5 to7 min aflatoxins can be separated using an Agilent XDB C18-column. We found that AFB1 and AFB2 were not found in D. denudatum. This was reliable through exceptionally low figures of fungal colonies observed after 6 hr of incubation. The developed analytical method is straightforward, be successfully used to determine the aflatoxins. Conclusion: The developed analytical method is straightforward, simple, accurate, economical and can be successfully used to find out the aflatoxins in remedial plants and consequently to have power over the quality of products. The presence of aflatoxin in the plant extracts was interrelated to the least fungal load in the remedial plants examined.

Keywords: aflatoxins, delphinium denudatum, liquid chromatography, mass spectrometry

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Authors: Jerman Madonsela, Wallace Matizamhuka, Akiko Yamamoto, Ronald Machaka, Brendon Shongwe

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In this study, biocompatibility evaluation of nanostructured near beta Ti-24Nb-4Zr-8Sn (Ti2448) alloy with non-toxic elements produced utilizing Spark plasma sintering (SPS) of very fine microsized powders attained through mechanical alloying was performed. The results were compared with pure titanium and Ti-6Al-4V (Ti64) alloy. Cell proliferation test was performed using murine osteoblastic cells, MC3T3-E1 at two cell densities; 400 and 4000 cells/mL for 7 days incubation. Pure titanium took a lead under both conditions suggesting that the presence of other oxide layers influence cell proliferation. No significant difference in cell proliferation was observed between Ti64 and Ti2448. Potentiodynamic measurement in Hanks, 0.9% NaCl and cell culture medium showed no distinct difference on the anodic polarization curves of the three alloys, indicating that the same anodic reaction occurred on their surface but with different rates. However, Ti2448 showed better corrosion resistance in cell culture medium with a slightly lower corrosion rate of 2.96 nA/cm2 compared to 4.86 nA/cm2 and 5.62 nA/cm2 of Ti and Ti64 respectively. Ti2448 adsorbed less protein as compared to Ti and Ti64 though no notable difference in surface wettability was observed.

Keywords: biocompatibility, osteoblast, corrosion, surface wettability, protein adsorption

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Authors: Michal Natan, Ori Gutman, Shlomo Margel, Ehud Banin

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Biofilm formation is a serious problem in medical and industrial settings due to the increased resistance of these communities to killing compared to free-living bacteria. This has prompted the search for agents that can inhibit both bacterial growth and biofilm formation. In this study, N-halamine rechargeable nanoparticles (NPs) were synthesized by co-polymerization of the monomer methacryl amide and the cross-linker monomer N,N-methylene bisacryl amide, and were subsequently loaded with Cl+, using bleach. The chlorinated NPs exhibited remarkable stability to organic reagents. The antibacterial mechanism of the P(MAA-MBAA)-Cl NPs involved generation of reactive oxygen species (ROS) only upon exposure to organic media, but not upon incubation in water, suggesting a specific activation. Moreover, a unique interaction of the P(MAA-MBAA)-Cl NPs with Staphylococcus aureus bacteria but not with human cells was discovered, whereby these microorganisms were all specifically targeted and marked for destruction. Finally, in collaboration with Netafim Ltd. irrigation drippers containing the P(MAA-MBAA)-Cl were incubated in the field and were shown to prevent fouling on them for 5 months as opposed to the control drippers that exhibited substantial fouling. Further, the NPs offer recharging to the surface, thus providing long-lasting protection that does not exist in the products available today. Taken together, the results demonstrate the great potential of implementing the charged NPs in devices and surfaces to prevent bacterial growth.

Keywords: bacteria, biofilm, fouling, nanoparticles

Procedia PDF Downloads 192

Authors: Ana M. Guzman, Claudia M. Rodriguez, Pedro F. B. Brandao, Elianna Castillo

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Cadmium (Cd) is a carcinogenic metal to which humans are exposed mainly due to its presence in the food chain. Lactic acid bacteria have the capability to bind cadmium and thus the potential to be used as probiotics to treat this metal toxicity in the human body. The main objective of this study is to evaluate the potential of native lactic acid bacteria, isolated from Colombian fermented cocoa, to remove cadmium from aqueous solutions. An initial screening was made with the Lactobacillus plantarum JCM 1055 type strain, and Cd was quantified by atomic absorption spectroscopy (AAS). Lb. plantarum JCM 1055 was grown in ½ MRS medium to follow growth kinetics during 32 h at 37 °C, by measuring optical density at 600 nm. Washed cells, grown for 18 h, were adjusted to obtain dry biomass concentrations of 1.5 g/L and 0.5 g/L for removal assays in 10 mL of Cd(NO₃)₂ solution with final concentrations of 10 mg/Kg or 1.0 mg/Kg. The assays were performed at two different pH values (2.0 and 5.0), and results showed better adsorption abilities at higher pH. After incubation for 1 h at 37 °C and 150 rpm, the removal percentages for 10 mg/Kg Cd with 1.5 g/L and 0.5 g/L biomass concentration at pH 5.0 were, respectively, 71% and 50%, while the efficiency was 9.15 and 4.52 mg Cd/g dry biomass, respectively. For the assay with 1.0 mg/Kg Cd at pH 5.0, the removal was 100% and 98%, respectively for the same biomass concentrations, and the efficiency was 1.63 and 0.56 mg Cd/g dry biomass, respectively. These results suggest the efficiency of Lactobacillus strains to remove cadmium and their potential to be used as probiotics to treat cadmium toxicity and reduce its accumulation in the human body.

Keywords: cadmium removal, fermented cocoa, lactic acid bacteria, probiotics

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