Search results for: sterile

Authors: Mohammad O. Diab, Sahera A. Saleh, Mustapha M. Dichari, Nijez Aloulou, Omar Hamoui, Feras Chehade

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This study sought to evaluate the efficiency of lead-free sterile radiation shield (Radionex) in the reduction of physician's exposure dose during interventional cardiology procedures. Cardiac catheterization procedures are often associated with high radiation doses and high levels of secondary radiation emitted by the patient's body. This study compares physician exposure dose rate during cardiac catheterization procedures done through the femoral artery with sterile radiation shielding to same procedures made without the shielding. The mean operator radiation dose rate without using the shield was found to be 18.4µSv/min compared to a mean dose rate of 5.1 µSv/min when using the shield, rendering a reduction of 72.5% of radiation received by the physician. Sterile radiation shielding is consequently an effective addition to a cardiac catheterization lab radiation protection system.

Keywords: cardiac catheterization, physician exposure dose, sterile radiation shielding, lead-free sterile radiation shields

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Authors: George Bibileishvili, Manana Mamulashvili, Zaza Javashvili, Liana Ebanoidze

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Economic Approaches to Obtaining and Maintaining Quality, Sterile Drinking Water from Natural Waters Through the Use of Nanotechnological Membrane Systems

Keywords: membrane, filter, ultrafiltration, water

Procedia PDF Downloads 46

Authors: Doris Lacej, Eni Bushi

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Cleaning and disinfection procedures are essential for maintaining the cleanliness status of the pharmaceutical manufacturing environment particularly of the cleanrooms and sterile unit area. The Good Manufacturing Practice (GMP) Annex 1 recommendation highly requires the implementation of the standard and validated cleaning and disinfection protocols. However, environmental monitoring has shown that even a validated cleaning method with certified agents may result in the presence of atypical microorganisms’ colony that exceeds GMP limits for a specific cleanroom area. In response to this issue, this case study aims to arrive at the root cause of the microbial contamination observed in the sterile production environment in Profarma pharmaceutical industry in Albania through applying a problem-solving practical approach that ensures the appropriate sterility grade. The guidelines and literature emphasize the importance of several factors in the prevention of possible microbial contamination occurring in cleanrooms, grade A and C. These factors are integrated into a practical framework, to identify the root cause of the presence of Aspergillus Niger colony in the sterile production environment in Profarma pharmaceutical industry in Albania. In addition, the application of a semi-automatic disinfecting system such as H2O2 FOG into sterile grade A and grade C cleanrooms has been an effective solution in eliminating the atypical colony of Aspergillus Niger. Selecting the appropriate detergents and disinfectants at the right concentration, frequency, and combination; the presence of updated and standardized guidelines for cleaning and disinfection as well as continuous training of operators on these practices in accordance with the updated GMP guidelines are some of the identified factors that influence the success of achieving sterility grade. However, to ensure environmental sustainability it is important to be prepared for identifying the source of contamination and making the appropriate decision. The proposed case-based practical approach may help pharmaceutical companies to achieve sterile production and cleanliness environmental sustainability in challenging situations. Apart from the integration of valid agents and standardized cleaning and disinfection protocols according to GMP Annex 1, pharmaceutical companies must be careful and investigate the source and all the steps that can influence the results of an abnormal situation. Subsequently apart from identifying the root cause it is important to solve the problem with a successful alternative approach.

Keywords: cleanrooms, disinfectants, environmental monitoring, GMP Annex 1

Procedia PDF Downloads 179

Authors: Fengmei Yao, Pengcheng Qin, Jiahua Zhang, Min Liu

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Global warming is expected to reduce the risk of low temperature stress in rice grown in temperate regions, but this impact has not been well verified by empirical studies directly on chilling injury in rice. In this study, a case study in Northeast China was presented to investigate whether the frequencies of chilling injury declined as a result of climate change, in comprehensive consideration of the potential effects from autonomous adaptation of rice production in response to climate change, such as shifts in cultivation timing and rice cultivars. It was found that frequency of total chilling injury (either delayed-growth type or sterile-type in a year) decreased but only to a limit extent in the context of climate change, mainly owing to a pronounced decrease in frequency of the delayed-growth chilling injury, while there was no overwhelming decreasing tendency for frequency of the sterile-type chilling injury, rather, it even increased considerably for some regions. If changes in cultivars had not occurred, risks of chilling injury of both types would have been much lower, specifically for the sterile-type chilling injury for avoiding deterioration in chilling sensitivity of rice cultivars. In addition, earlier planting helped lower the risk of chilling injury but still can not overweight the effects of introduction of new cultivars. It was concluded that risks of chilling injury in rice would not necessarily decrease as a result of climate change, considering the accompanying adaptation process may increase the chilling sensitivity of rice production system in a warmer climate conditions, and thus precautions should still be taken.

Keywords: chilling injury, rice, CERES-rice model, climate warming, North east China

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Authors: Abdullahi Mohammed

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The microbial quality of beef and mutton sold in four major markets of Bauchi metropolis was assessed in order to assist in ascertaining safety. Shops were selected from 'Muda Lawal', 'Yelwa', 'Wunti', and 'Gwallameji' markets. The total bacterial count was used as index of quality. A total of thirty two (32) samples were collected in two successive visits. The samples were packed and labelled in a sterile polythene bags for transportation to the laboratory. Microbial analysis was carried out immediately upon arrival under a septic condition, where aerobic plate was used in determining the microbial load. Result showed that beef and mutton from Gwallameji had the highest bacterial count of 9.065 X 105 cfu/ml and 8.325 X 105 cfu/ml for beef and mutton respectively followed by Wunti market (6.95 X 105 beef and 4.838 X 105 motton) and Muda Lawal (4.86 X 105 cfu/ml beef and 5.998 X 105 cfu/ml mutton). Yelwa had 5.175 X 105 and 5.30 X 105 for beef and mutton respectively. Bacterial species isolated from the samples were Escherichia coli, Salmonella spp, Streptococcus species and Staphylococcus species. However, results obtained from all markets showed that there was no significant differences between beef and mutton in terms of microbial quality.

Keywords: beef, mutton, salmonella, sterile

Procedia PDF Downloads 418

Authors: Abul B. M. M. K. Islam, Mandar Dave, Roderick V. Jensen, Ashok R. Amin

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A systems approach was applied to characterize differentially expressed transcripts, bioinformatics pathways, and proteins and prostaglandins (PGs) from lung fibroblasts procured from wild-type (WT), COX-1-/- and COX-2-/- mice to understand system level control mechanism. Bioinformatics analysis of COX-2 and COX-1 ablated cells induced COX-1 and COX-2 specific signature respectively, which significantly overlapped with an 'IL-1β induced inflammatory signature'. This defined novel cross-talk signals that orchestrated coordinated activation of pathways of sterile inflammation sensed by cellular stress. The overlapping signals showed significant over-representation of shared pathways for interferon y and immune responses, T cell functions, NOD, and toll-like receptor signaling. Gene Ontology Biological Process (GOBP) and pathway enrichment analysis specifically showed an increase in mRNA expression associated with: (a) organ development and homeostasis in COX-1-/- cells and (b) oxidative stress and response, spliceosomes and proteasomes activity, mTOR and p53 signaling in COX-2-/- cells. COX-1 and COX-2 showed signs of functional pathways committed to cell cycle and DNA replication at the genomics level. As compared to WT, metabolomics analysis revealed a significant increase in COX-1 mRNA and synthesis of basal levels of eicosanoids (PGE2, PGD2, TXB2, LTB4, PGF1α, and PGF2α) in COX-2 ablated cells and increase in synthesis of PGE2, and PGF1α in COX-1 null cells. There was a compensation of PGE2 and PGF1α in COX-1-/- and COX-2-/- cells. Collectively, these results support a broader, differential and collaborative regulation of both COX-1 and COX-2 pathways at the metabolic, signaling, and genomics levels in cellular homeostasis and sterile inflammation induced by cellular stress.

Keywords: cyclooxygenases, inflammation, lung fibroblasts, systemic

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Authors: Escada A. L. A., Pereira C. A., Jorge A. O. C., Alves Claro A. P. R.

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In the present work, a susceptibility and efficacy of the Ti–7.5Mo alloy nanotube and Ti–7.5Mo alloy to bacterial biofilm formation after surface treatment was evaluated. The Ti–7.5Mo alloy was obtained in arc furnace under an argon atmosphere. Ingots were then homogenized under vacuum at 1100 ◦C for 86.4 ks to eliminate chemical segregation and after cold worked discs were cutting. Nanotubes were processed using anodic oxidation in 0.25% NH4F electrolyte solution. Biofilms were grown in discs immersed in sterile brain heart infusion broth (BHI) containing 5% sucrose, inoculated with microbial suspension (106 cells/ml) and incubated for 5 days. Next, the discs were placed in tubes with sterile physiological solution 0.9% sodium chloride (NaCl) and sonicated for to disperse the biofilms. Tenfold serial dilutions were carried and aliquots seeded in selective agar, which were then incubated for 48 h. Then, the numbers CFU/ml (log 10) were counted and analyzed statistically. Scanning electron microscopy (SEM) on discs with biofilms groupswas performed, atomic force microscope (AFM) and contact angle. The results show that there is no difference in bacterial adhesion between Ti–7.5Mo alloy nanotube pure titanium and Ti–7.5Mo alloy.

Keywords: biofilm, titanium alloy, brain heart infusion, scanning electron microscopy

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Authors: Mehrdad Ahmadi, Mohamad Babaie, Shiva Osouli, Bahareh Salehi, Nadia Kalantaraian

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The olive fruit fly, Bactrocera oleae Gmelin (Diptera: Tephritidae), is one of the most serious pests in olive orchards in growing province in Iran. The female lay eggs in green olive fruit and larvae hatch inside the fruit, where they feed upon the fruit matters. One of the main ecologically friendly and species-specific systems of pest control is the sterile insect technique (SIT) which is based on the release of large numbers of sterilized insects. The objective of our work was to develop a SIT against B. oleae by using of gamma radiation for the laboratory and field trial in Iran. Oviposition of female mated by irradiated males is one of the main parameters to determine achievement of SIT. To conclude the sterile dose, pupae were placed under 0 to 160 Gy of gamma radiation. The main factor in SIT is the productivity of females which are mated by irradiated males. The emerged adults from irradiated pupae were mated with untreated adults of the same age by confining them inside the transparent cages. The fecundity of the irradiated males mated with non-irradiated females was decreased with the increasing radiation dose level. It was observed that the number of eggs and also the percentage of the egg hatching was significantly (P < 0.05) affected in either IM x NF crosses compared with NM x NF crosses in F₁ generation at all doses. Also, the statistical analysis showed a significant difference (P < 0.05) in the mean number of eggs laid between irradiated and non-irradiated females crossed with irradiated males, which suggests that the males were susceptible to gamma radiation. The egg hatching percentage declined markedly with the increase of the radiation dose of the treated males in mating trials which demonstrated that egg hatch rate was dose dependent. Our results specified that gamma radiation affects the longevity of irradiated B. oleae larvae (established from irradiated pupae) and significantly increased their larval duration. Results show the gamma radiation, and SIT can be used successfully against olive fruit flies.

Keywords: fertility, olive fruit fly, radiation, sterile insect technique

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Authors: Hasan AL-Khshemawee, Manjree Agarwal, Xin Du, Yonglin Ren

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The possibility use of stable isotopes to study Medfly mating and life history were investigated in these experiments. 13C6 glucose was incorporated in the diet of the Mediterranean fruit fly Ceratitis capitata (Diptera: Tephidae). Treatments included labelling and unlabelled of either the media or adult sugar water. The measured started from egg hatching till the adults have died. After mating, the adults were analysed for 13C6 glucose ratio using Liquid chromatography-mass spectrometry LC-MS in two periods of time immediately and after three days of mating. Results showed that stable isotopes were used successfully for labelling Medfly in laboratory conditions, and there were significant differences between labelled and unlabelled treatment in eggs hatching, larval development, pupae emergence, survival of adults and mating behaviour. Labelling during larval development and combined labelling of larvae and adults resulted in detectable values. The label glucose in larvae stage did not effect on mating behaviour, however, the label glucose in adults’ stage was affected by mating behaviour. We recommended that it is possible to label adults of Mediterranean fruit fly C. capitata and detected the label after mating. This method offers good tools to study mating behaviour in Medfly and other types of insects and could be providing useful tools in genetic studies, sterile insect technique (SIT) or agricultural pest management. Also, we recommended using this technique in the field.

Keywords: stable isotope, sterile insect technique (SIT), medfly, mating behaviour

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Authors: Sharon K. Mahlangu, Mustapha Mohammed, Felix D. Dakora

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Rural households in Africa depend largely on legumes as a source of high-protein food due to N₂-fixation by rhizobia when they infect plant roots. However, the legume/rhizobia symbiosis can exhibit some level of specificity such that some legumes may be selectively nodulated by only a particular group of rhizobia. In contrast, some legumes are highly promiscuous and are nodulated by a wide range of rhizobia. Little is known about the nodulation promiscuity of bacterial symbionts from wild legumes such as Aspalathus linearis, especially if they can nodulate cultivated grain legumes such as cowpea and Kersting’s groundnut. Determining the host range of the symbionts of wild legumes can potentially reveal novel rhizobial strains that can be used to increase nitrogen fixation in cultivated legumes. In this study, bacteria were isolated and tested for their ability to induce root nodules on their homologous hosts. Seeds were surface-sterilized with alcohol and sodium hypochlorite and planted in sterile sand contained in plastic pots. The pot surface was covered with sterile non-absorbent cotton wool to avoid contamination. The plants were watered with nitrogen-free nutrient solution and sterile water in alternation. Three replicate pots were used per isolate. The plants were grown for 90 days in a naturally-lit glasshouse and assessed for nodulation (nodule number and nodule biomass) and shoot biomass. Seven isolates from each of Kersting’s groundnut and cowpea and two from Rooibos tea plants were tested for their ability to nodulate soybean, mung bean, common bean and Bambara groundnut. The results showed that of the isolates from cowpea, where VUSA55 and VUSA42 could nodulate all test host plants, followed by VUSA48 which nodulated cowpea, Bambara groundnut and soybean. The two isolates from Rooibos tea plants nodulated Bambara groundnut, soybean and common bean. However, isolate L1R3.3.1 also nodulated mung bean. There was a greater accumulation of shoot biomass when cowpea isolate VUSA55 nodulated common bean. Isolate VUSA55 produced the highest shoot biomass, followed by VUSA42 and VUSA48. The two Kersting’s groundnut isolates, MGSA131 and MGSA110, accumulated average shoot biomass. In contrast, the two Rooibos tea isolates induced a higher accumulation of biomass in Bambara groundnut, followed by common bean. The results suggest that inoculating these agriculturally important grain legumes with cowpea isolates can contribute to improved soil fertility, especially soil nitrogen levels.

Keywords: legumes, nitrogen fixation, nodulation, rhizobia

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Authors: Nurul Aliaa Idris, David A. Collings

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Phi thickenings are specialized secondary cell wall thickenings that are found in the cortex of the roots in a wide range of plant species, including orchids. The role of phi thickenings in the root is still under debate through research have linked environmental conditions, particularly abiotic stresses such as water stress, heavy metal stress and salinity to their induction in the roots. It has also been suggested that phi thickenings may act as a barrier to regulate solute uptake, act as a physical barrier against fungal hyphal penetration due to its resemblance to the Casparian strip and play a mechanical role to support cortical cells. We have investigated phi thickening function in epiphytic orchids of the genus Miltoniopsis through induction experiment against factors such as soil compaction and water stress. The permeability of the phi thickenings in Miltoniopsis was tested through uptake experiments using the fluorescent tracer dyes Calcofluor white, Lucifer yellow and Propidium iodide then viewed with wide-field or confocal microscopy. To test whether phi thickening may prevent fungal colonization in the root cell, fungal re-infection experiment was conducted by inoculating isolated symbiotic fungus to sterile in vitro Miltoniopsis explants. As the movement of fluorescent tracers through the apoplast was not blocked by phi thickenings, and as phi thickenings developed in the roots of sterile cultures in the absence of fungus and did not prevent fungal colonization of cortical cells, the phi thickenings in Miltoniopsis do not function as a barrier. Phi thickenings were found to be absent in roots grown on agar and remained absent when plants were transplanted to moist soil. However, phi thickenings were induced when plants were transplanted to well-drained media, and by the application of water stress in all soils tested. It is likely that phi thickenings stabilize the root cortex during dehydration. Nevertheless, the varied induction responses present in different plant species suggest that the phi thickenings may play several adaptive roles, instead of just one, depending on species.

Keywords: abiotic stress, Miltoniopsis, orchid, phi thickening

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Authors: Ksenija Taški-Ajdukovic, Nevena Nagl, Živko Ćurčić, Dario Danojević

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Information about genetic diversity of sugar beet parental populations is of a great importance for hybrid breeding programs. The aim of this research was to evaluate genetic diversity among and within populations and lines of diploid sugar beet pollinators, by using SSR markers. As plant material were used eight pollinators originating from three USDA-ARS breeding programs and four pollinators from Institute of Field and Vegetable Crops, Novi Sad. Depending on the presence of self-fertility gene, the pollinators were divided into three groups: autofertile (inbred lines), autosterile (open-pollinating populations), and group with partial presence of autofertility gene. A total of 40 SSR primers were screened, out of which 34 were selected for the analysis of genetic diversity. A total of 129 different alleles were obtained with mean value 3.2 alleles per SSR primer. According to the results of genetic variability assessment the number and percentage of polymorphic loci was the maximal in pollinators NS1 and tester cms2 while effective number of alleles, expected heterozygosis and Shannon’s index was highest in pollinator EL0204. Analysis of molecular variance (AMOVA) showed that 77.34% of the total genetic variation was attributed to intra-varietal variance. Correspondence analysis results were very similar to grouping by neighbor-joining algorithm. Number of groups was smaller by one, because correspondence analysis merged IFVCNS pollinators with CZ25 into one group. Pollinators FC220, FC221 and C 51 were in the next group, while self-fertile pollinators CR10 and C930-35 from USDA-Salinas were separated. On another branch were self-sterile pollinators ЕL0204 and ЕL53 from USDA-East Lansing. Sterile testers cms1 and cms2 formed separate group. The presented results confirmed that SSR analysis can be successfully used in estimation of genetic diversity within and among sugar beet populations. Since the tested pollinator differed considering the presence of self-fertility gene, their heterozygosity differed as well. It was lower in genotypes with fixed self-fertility genes. Since the most of tested populations were open-pollinated, which rarely self-pollinate, high variability within the populations was expected. Cluster analysis grouped populations according to their origin.

Keywords: auto fertility, genetic diversity, pollinator, SSR, sugar beet

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Authors: Ofonime Ogba, Augustine Akpan

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Objectives and Goals: The study was aimed at determining pathogenic microbial carriage on the external body parts of Camponotus consobrinus which is also known as the banded sugar ant because of its liking for sugar and sweet food. The level of pathogenic microbial carriage of Camponotus consobrinus in association to the environment in which they have been collected is not known. Methods: The ants were purposively collected from four locations including the kitchens, bedroom of various homes, food shops, and bakeries. The sample collection took place within the hours of 6:30 pm to 11:00 pm. The ants were trapped in transparent plastic containers of which sugar, pineapple peels, sugar cane and soft drinks were used as bait. The ants were removed with a sterile spatula and put in 10mls of peptone water in sterile universal bottles. The containers were vigorously shaken to wash the external surface of the ant. It was left overnight and transported to the Microbiology Laboratory, University of Calabar Teaching Hospital for analysis. The overnight peptone broths were inoculated on Chocolate agar, Blood agar, Cystine Lactose Electrolyte-Deficient agar (CLED) and Sabouraud dextrose agar. Incubation was done aerobically and in a carbon dioxide jar for 24 to 48 hours at 37°C. Isolates were identified based on colonial characteristics, Gram staining, and biochemical tests. Results: Out of the 250 Camponotus consobrinus caught for the study, 90(36.0%) were caught in the kitchen, 75(30.0%) in the bedrooms 40(16.0%) in the bakery while 45(18.0%) were caught in the shops. A total of 82.0% prevalence of different microbial isolates was associated with the ants. The kitchen had the highest number of isolates 75(36.6%) followed by the bedroom 55(26.8%) while the bakery recorded the lowest number of isolates 35(17.1%). The profile of micro-organisms associated with Camponotus consobrinus was Escherichia coli 73(30.0%), Morganella morganii 45(18.0%), Candida species 25(10.0%), Serratia marcescens 10(4.0%) and Citrobacter freundii 10(4.0%). Conclusion: Most of the Camponotus consobrinus examined in the four locations harboured potential pathogens. The presence of ants in homes and shops can facilitate the propagation and spread of pathogenic microorganisms. Therefore, the development of basic preventive measures and the control of ants must be taken seriously.

Keywords: Camponotus consobrinus, potential pathogens, microbial isolates, spread

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Authors: Abhay Asthana, Gyati Shilakari Asthana

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It’s the patient compliance and stability in combination with controlled drug delivery and biocompatibility that forms the core feature in present research and development of sustained biodegradable patch formulation intended for wound healing. The aim was to impart sustained degradation, sterile formulation, significant folding endurance, elasticity, biodegradability, bio-acceptability and strength. The optimized formulation was developed using component including polymers including Hydroxypropyl methyl cellulose, Ethylcellulose, and Gelatin, and Citric Acid PEG Citric acid (CPEGC) triblock dendrimers and active Curcumin. Polymeric mixture dissolved in geometric order in suitable medium through continuous stirring under ambient conditions. With continued stirring Curcumin was added with aid of DCM and Methanol in optimized ratio to get homogenous dispersion. The dispersion was sonicated with optimum frequency and for given time and later casted to form a patch form. All steps were carried out under under strict aseptic conditions. The formulations obtained in the acceptable working range were decided based on thickness, uniformity of drug content, smooth texture and flexibility and brittleness. The patch kept on stability using butter paper in sterile pack displayed folding endurance in range of 20 to 23 times without any evidence of crack in an optimized formulation at room temperature (RT) (24 ± 2°C). The patch displayed acceptable parameters after stability study conducted in refrigerated conditions (8±0.2°C) and at RT (24 ± 2°C) upto 90 days. Further, no significant changes were observed in critical parameters such as elasticity, biodegradability, drug release and drug content during stability study conducted at RT 24±2°C for 45 and 90 days. The drug content was in range 95 to 102%, moisture content didn’t exceeded 19.2% and patch passed the content uniformity test. Percentage cumulative drug release was found to be 80% in 12h and matched the biodegradation rate as drug release with correlation factor R2>0.9. The biodegradable patch based formulation developed shows promising results in terms of stability and release profiles.

Keywords: sustained biodegradation, wound healing, polymers, stability

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Authors: Paulina Zavistanaviciute, Vita Krungleviciute, Elena Bartkiene

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Lactic acid bacteria (LAB) are microbial supplements that increase the nutritional, therapeutic, and safety value of food and feed. Often LAB strains are incubated in an expensive commercially available de Man-Rogosa-Sharpe (MRS) medium; the cultures are centrifuged, and the cells are washing with sterile water. Potato juice and apple juice industry bio-products are industrial wastes which may constitute a source of digestible nutrients for microorganisms. Due to their low cost and good chemical composition, potato juice and apple juice production bio- products could have a potential application in LAB encapsulation. In this study, pure LAB (P. acidilactici and P. pentosaceus) were multiplied in a crushed potato juice and apple juice industry bio-products medium. Before using, bio-products were sterilized and filtered. No additives were added to mass, except apple juice industry bioproducts were diluted with sterile water (1/5; v/v). The tap of sterilised mass, and LAB cell suspension (5 mL), containing of 8.9 log10 colony-forming units (cfu) per mL of the P. acidilactici and P. pentosaceus was used to multiply the LAB for 72 h. The final colony number in the potato juice and apple juice bio- products substrate was on average 9.60 log10 cfu/g. In order to stabilize the LAB, several methods of dehydration have been tested: lyophilisation (MilrockKieffer Lane, Kingston, USA) and dehydration in spray drying system (SD-06, Keison, Great Britain). Into the spray drying system multiplied LAB in a crushed potato juice and apple juice bio-products medium was injected in peristaltic way (inlet temperature +60 °C, inlet air temperature +150° C, outgoing air temperature +80 °C, air flow 200 m3/h). After lyophilisation (-48 °C) and spray drying (+150 °C) the viable cell concentration in the fermented potato juice powder was 9.18 ± 0.09 log10 cfu/g and 9.04 ± 0.07 log10 cfu/g, respectively, and in apple mass powder 8.03 ± 0.04 log10 cfu/g and 7.03 ± 0.03 log10 cfu/g, respectively. Results indicated that during the storage (after 12 months) at room temperature (22 +/- 2 ºC) LAB count in dehydrated products was 5.18 log10 cfu/g and 7.00 log10 cfu/g (in spray dried and lyophilized potato juice powder, respectively), and 3.05 log10 cfu/g and 4.10 log10 cfu/g (in spray dried and lyophilized apple juice industry bio-products powder, respectively). According to obtained results, potato juice could be used as alternative substrate for P. acidilactici and P. pentosaceus cultivation, and by drying received powders can be used in food/feed industry as the LAB starters. Therefore, apple juice industry by- products before spray drying and lyophilisation should be modified (i. e. by using different starches) in order to improve its encapsulation.

Keywords: bio-products, encapsulation, lactic acid bacteria, sustainability

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Authors: Indu Gaur, Neha Bhadauria, Shilpi Shilpi, Susmita Goswami, Prem D. Sharma, Prabir K. Paul

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The concept of organic agriculture has been accepted as novelty in Indian society, but there is no data available on the human pathogens colonizing plant parts due to such practices. Also, the pattern and mechanism of their colonization need to be understood in order to devise possible strategies for their prevention. In the present study, human pathogenic bacteria were isolated from organically grown tomato plants and five of them were identified as Klebsiella pneumoniae, Enterobacter ludwigii, Serratia fonticola, Stenotrophomonas maltophilia and Chryseobacterium jejuense. Tomato plants were grown in controlled aseptic conditions with 25±1˚C, 70% humidity and 12 hour L/D photoperiod. Six weeks old plants were divided into 6 groups of 25 plants each and treated as follows: Group 1: K. pneumonia, Group 2: E. ludwigii, Group 3: S. fonticola, Group 4: S. maltophilia, Group 5: C. jejuense, Group 6: Sterile distilled water (control). The inoculums for all treatments were prepared by overnight growth with uniform concentration of 108 cells/ml. Leaf samples from above groups were collected at 0.5, 2, 4, 6 and 24 hours post inoculation for the colony forming unit counts (CFU/cm2 of leaf area) of individual pathogens using leaf impression method. These CFU counts were used for the in vivo colonization assay and adherence assay of individual pathogens. Also, resistance of these pathogens to at least 12 antibiotics was studied. Based on these findings S. fonticola was found to be most prominently colonizing the phylloplane of tomato and was further studied. Tomato plants grown in controlled aseptic conditions same as mentioned above were divided into 2 groups of 25 plants each and treated as follows: Group 1: S. fonticola, Group 2: Sterile distilled water (control). Leaf samples from above groups were collected at 0, 24, 48, 72 and 96 hours post inoculation and homogenized in suitable buffers for surface and cell wall protein isolation. Protein samples thus obtained were subjected to isocratic SDS-gel electrophoresis and analyzed. It was observed that presence of S. fonticola could induce the expression of at least 3 additional cell wall proteins at different time intervals. Surface proteins also showed variation in the expression pattern at different sampling intervals. Further identification of these proteins by MALDI-MS and bioinformatics tools revealed the gene(s) involved in the interaction of S. fonticola with tomato phylloplane.

Keywords: cell wall proteins, human pathogenic bacteria, phylloplane, solanum lycopersicum

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Authors: Elisabetta Cianfanelli, Margherita Tufarelli, Paolo Pupparo

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The dialectical mediation between desires and objects or between mass production and consumption continues to evolve over time. This relationship is influenced both by variable geometries of contexts that are distant from the mere design of product form and by aspects rooted in the very definition of industrial design. In particular, the overcoming of macro-areas of innovation in the technological, social, cultural, formal, and morphological spheres, supported by recent theories in critical and speculative design, seems to be moving further and further away from the design of the formal dimension of advanced products. The articulated fabric of theories and practices that feed the definition of “hyperobjects”, and no longer objects describes a common tension in all areas of design and production of industrial products. The latter are increasingly detached from the design of the form and meaning of the same in mass productions, thus losing the quality of products capable of social transformation. For years we have been living in a transformative moment as regards the design process in the definition of the industrial product. We are faced with a dichotomy in which there is, on the one hand, a reactionary aversion to the new techniques of industrial production and, on the other hand, a sterile adoption of the techniques of mass production that we can now consider traditional. This ambiguity becomes even more evident when we talk about industrial products, and we realize that we are moving further and further away from the concepts of "form" as a synthesis of a design thought aimed at the aesthetic-emotional component as well as the functional one. The design of forms and their contents, as statutes of social acts, allows us to investigate the tension on mass production that crosses seasons, trends, technicalities, and sterile determinisms. The design culture has always determined the formal qualities of objects as a sum of aesthetic characteristics functional and structural relationships that define a product as a coherent unit. The contribution proposes a reflection and a series of practical experiences of research on the form of advanced products. This form is understood as a kaleidoscope of relationships through the search for an identity, the desire for democratization, and between these two, the exploration of the aesthetic factor. The study of form also corresponds to the study of production processes, technological innovations, the definition of standards, distribution, advertising, the vicissitudes of taste and lifestyles. Specifically, we will investigate how the genesis of new forms for new meanings introduces a change in the relative innovative production techniques. It becomes, therefore, fundamental to investigate, through the reflections and the case studies exposed inside the contribution, also the new techniques of production and elaboration of the forms of the products, as new immanent and determining element inside the planning process.

Keywords: industrial design, product advanced design, mass productions, new meanings

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Authors: F. Pal-Fam, R. Hoffmann, S. Keszthelyi

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Granary weevil, Sitophilus granarius (L.) (Col.: Curculionidae) is a typical cosmopolitan pest. It can cause significant damage to stored grains, and can drastically decrease yields. Damaged grain has reduced nutritional and market value, weaker germination, and reduced weight. The commonly used protectants against stored-product pests in Europe are residual insecticides, applied directly to the product. Unfortunately, these pesticides can be toxic to mammals, the residues can accumulate in the treated products, and many pest species could become resistant to the protectants. During recent years, alternative solutions of grain protection have received increased attention. These solutions are considered as the most promising alternatives to residual insecticides. The aims of our comparative study were to obtain information about the efficacies of the 1. diatomaceous earth, 2. sterile insect technology and 3. herbal oils against the S. granarius on grain (foremost maize), and to evaluate the influence of the dose rate on weevil mortality and progeny. The main results of our laboratory experiments are the followings: 1. Diatomaceous earth was especially efficacious against S. granarius, but its insecticidal properties depend on exposure time and applied dose. The efficacy on barley was better than on maize. Mortality value of the highest dose was 85% on the 21st day in the case of barley. It can be ascertained that complete elimination of progeny was evidenced on both gain types. To summarize, a satisfactory efficacy level was obtained only on barley at a rate of 4g/kg. Alteration of efficacy between grain types can be explained with differences in grain surface. 2. The mortality consequences of Roentgen irradiation on the S. granarius was highly influenced by the exposure time, and the dose applied. At doses of 50 and 70Gy, the efficacy accepted in plant protection (mortality: 95%) was recorded only on the 21st day. During the application of 100 and 200Gy doses, high mortality values (83.5% and 97.5%) were observed on the 14th day. Our results confirmed the complete sterilizing effect of the doses of 70Gy and above. The autocide effect of 50 and 70Gy doses were demonstrated when irradiated specimens were mixed into groups of fertile specimens. Consequently, these doses might be successfully applied to put sterile insect technique (SIT) into practice. 3. The results revealed that both studied essential oils (Callendula officinalis, Hippophae rhamnoides) exerted strong toxic effect on S. granarius, but C. officinalis triggered higher mortality. The efficacy (94.62 ± 2.63%) was reached after a 48 hours exposure to H. rhamnoides oil at 2ml/kg while the application of 2ml/kg of C. officinalis oil for 24 hours produced 98.94 ± 1.00% mortality rate. Mortality was 100% at 5 ml/kg of H. rhamnoides after 24 hours duration of its application, while with C. officinalis the same value could be reached after a 12 hour-exposure to the oil. Both essential oils applied were eliminated the progeny.

Keywords: Sitophilus granarius, stored product, protection, alternative solutions

Procedia PDF Downloads 143

Authors: Ayesha Irum, Sadia Mumtaz, Abdul Rehman, Iffat Naz, Safia Ahmed

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The present study was conducted to evaluate the potential applicability of biological trickling filter system for the treatment of simulated textile wastewater containing reactive azo dyes with bacterial consortium under non-sterile conditions. The percentage decolorization for the treatment of wastewater containing structurally different dyes was found to be higher than 95% in all trials. The stable bacterial count of the biofilm on stone media of the trickling filter during the treatment confirmed the presence, proliferation, dominance and involvement of the added microbial consortium in the treatment of textile wastewater. Results of physicochemical parameters revealed the reduction in chemical oxygen demand (58.5-75.1%), sulphates (18.9-36.5%), and phosphates (63.6-73.0%). UV-Visible and FTIR spectroscopy confirmed decolorization of dye containing wastewater was the ultimate consequence of biodegradation. Toxicological studies revealed the nontoxic nature of degradative metabolites.

Keywords: biodegradation, textile dyes, waste water, trickling filters

Procedia PDF Downloads 396

Authors: Sridevi Devadas

Abstract:

This study was aimed to examine the combination efficiency of Clorox (5.25% Sodium Hypochlorite) and mercury chloride (HgCl2) as reagent for surface sterilization process of aquatic plant, Cryptocoryne affinis (C. affinis). The treatment applied 10% of the Clorox and 0.1 ppm of mercury chloride. The maximum exposure time for Clorox and mercury chloride was 10 min and 60 sec respectively. After exposed to the treatments protocols (T1-T15) the explants were transferred to culture room under control temperature at 25°C ± 2°C and subjected to 16 hours fluorescence light (2000 lumens) for 30 days. The both sterilizing agents were not applied on control specimens. Upon analysis, the result indicates all of the treatments protocols produced sterile explants at range of minimum 1.5 ± 0.7 (30%) to maximum 5.0 ± 0.0 (100%). Meanwhile, maximum 1.0 ± 0.7 numbers of leaves and 1.4 ± 0.6 numbers of roots have been produced. The optimized exposure time was 0 to 15 min for Clorox and 30 sec for HgCl2 whereby 90% to 100% sterilization was archived at this condition.

Keywords: Cryptocoryne affinis, surface sterilization, tissue culture, clorox, mercury chloride

Procedia PDF Downloads 560

Authors: Sisuwan Kaseamsawat, Sivapan Choo-In

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This study aimed to evaluated the surface water quality for agriculture and consumption in the district. Surface water quality parameters in this study in cluding water temperature, turbidity, conductivity. salinity, pH, dissolved oxygen, BOD, nitrate, Suspended solids, phosphorus. Total dissolve solids, iron, copper, zinc, manganese, lead and cadmium. Water samples were collected from small excavation, Lychee, Pomelo, and Coconut orchard for 3 season during January to December 2011. The surface water quality from small excavation, Lychee, pomelo, and coconut orchard are meet the type III of surface water quality standard issued by the National Environmental Quality Act B. E. 1992. except the concentration of heavy metal. And did not differ significantly at 0.05 level, except dissolved oxygen. The water is suitable for consumption by the usual sterile and generally improving water quality through the process before. And is suitable for agriculture.

Keywords: water quality, surface water quality, Thailand, water

Procedia PDF Downloads 325

Authors: Sridevi Devadas

Abstract:

This study was aimed to examine the combination efficiency of Clorox (5.25% Sodium Hypochlorite) and mercury chloride (HgCl2) as a reagent for surface sterilization process of aquatic plant and cryptocoryne affinis (C. affinis). The treatment applied 10% of the Clorox and 0.1ppm of mercury chloride. The maximum exposure time for clorox and mercury chloride was 10min and 60sec respectively. After exposed to the treatments protocols (T1-T15) the explants were transferred to culture room under control temperature at 25°C ± 2°C and subjected to 16 hours fluorescence light (2000 lumens) for 30 days. The both sterilizing agents were not applied on control specimens. Upon analysis, The result indicates all of the treatments protocols produced sterile explants at range of minimum 1.5 ± 0.7 (30%) to maximum 5.0 ± 0.0 (100%). Meanwhile, maximum 1.0 ± 0.7 numbers of leaves and 1.4 ± 0.6 numbers of roots have been produced. The optimized exposure time was 0 to 15 min for Clorox and 30 sec for HgCl2 whereby 90% to 100% sterilization was archived at this condition.

Keywords: Cryptocoryne affinis, surface sterilization, tissue culture, clorox, mercury chloride

Procedia PDF Downloads 343

Authors: Elham Rouhollahi, Soheil Zorofchian Moghadamtousi, Salma Baig, Mahmood Ameen Abdulla, Zahurin Mohamed

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This study was designed to assess cutaneous wound healing potential of hexane extract of Curcuma purpurascens rhizomes (HECP). Twenty-four rats were divided into 4 groups: 1. Negative, 2. Low dose, 3. High dose and 4. Treatment, with 6 rats in each group. Full-thickness incisions with a diameter of 2 cm were made on the back of each rat. Rats were topically treated two times a day for 15 days. Group 1-4 were treated with sterile distilled water, 5% and 10% of extract and intrasite gel, respectively. Masson's trichrome and hematoxylin staining techniques are employed for histological analysis revealed strong wound healing potential closer to that of conventional drug intrasite gel. HECP significantly decreased wound area and an increase in hydroxyproline, cellular proliferation, the number of blood vessels and the level of collagen synthesis was observed. Thus, it could be concluded that HECP possesses strong wound healing potential.

Keywords: Curcuma purpurascens, wound healing, histopathology, hematoxylin staining

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Authors: Zahra Tabarsi, Mehdi Marjani, Alireza Najafpour, Alborz Mirzade

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The aim of this study was to evaluate and compare the macroscopic effect of low level laser and plasma jet for wound healing in rats. The study was performed on 40 old male white rats with an average weight of 250 g and an average age of the same age. After preparing the rats from Ibn Sina Research Institute, they were kept the same for one week under environmental conditions such as temperature, humidity and light, and nutrition such as the type of diet and the number of meals. Then, to start the research, rats were randomly divided into two groups (A): laser treatment of wounds, group (B): plasma wound treatment. All rats were inhibited 4 hours before each anesthesia under conditions of abstinence and up to 2 hours after drinking water. Rats were anesthetized by intraperitoneal injection of ketamine 10% and xylazine 2%.After scrubbing between two shoulders of each rat, a circular wound was created by sterile 5 mm biopsy puncture. Group A rats were treated with low level laser in three sessions and group B in three sessions with argon plasma. Based on the observed results, it seems that Low level laser radiation has more acceptable and appropriate effects than cold plasma on the healing of rat skin wounds.

Keywords: low-level laser, plasma jet, rat, wound healing

Procedia PDF Downloads 78

Authors: Liu Yuhong, Hussen Mansai, Mei Chunli

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Inflammatory discitis is a sterile inflammatary disease that typically presents with abnormalities in two adjacent vertebral bodies and the intervening disk. Diagnosis this disorder is usually difficult and ideal management remains controversial. In this report，we examine a case of inflammatory discitis in a 56 year old female in which treatment with adalimumab ameliorated symptoms. The 56-year-old female patient developed repeatedly inflammatory discitis in the past three years, presenting with severe back pain, an elevated C-reactive protein and erythrocyte sedimentation rate, radiological erosive changes in vertebral and intervertebral disk of the spine. Surgical treatment, antibiotics and non steroidal anti-inflammatory drugs(NSAIDs) were used, but the patient still suffered from recurrent onset of unbearable backache. Three years later from the patient’s first admission，adalimumab was prescribed due to the third occurrence of Anderson lesions, which she had been suffering from for years. Soon after the same day of adalimumab therapy, her symptoms had a dramatic improvement. On the following day she could stand and walk slowly, her CRP and ESR were decreased to nearly normal levels in 4 weeks. Human leukocyte antigen (HLA)-typing analysis revealed a positive result for HLA-B27, the patient’s inflammatory discitis was considered to be associated with spondyloarthropathy.

Keywords: adalimumab, inflammatory discitis, spondyloarthropathy, patient

Procedia PDF Downloads 225

Authors: I-En Lin, Feng-Jung Yang

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In addition to kidney transplant, renal replacement therapy involves hemodialysis and peritoneal dialysis (PD). PD possesses advantages such as maintaining stable physiological blood status and blood pressure, alleviating anemia, and improving mobility, which make it an ideal method for at-home dialysis treatment. However, potential danger still exists despite the numerous advantages of PD, particularly when patients require dialysis exchange four to five times a day, during which improper operation can easily lead to peritonitis. The process of draining and filling is called an exchange and takes about 30 to 40 minutes. Connecting the transfer set requires sterile technique. Transfer set may require a new cap each time that it disconnects from the bag after an exchange. There are many chances to get infection due to unsafe behavior (ex: hand tremor, poor eyesight and weakness, cap fall-down). The proposed semi-automatic connection mechanism used in the PD can greatly reduce infection chances. This light-weight connection device is portable. The device also does not require using throughout the entire process. It is capable of significantly improving quality of life. Therefore, it is very promising to adopt in home care application.

Keywords: automatic connection, catheter, glomerulonephritis, peritoneal dialysis

Procedia PDF Downloads 206

Authors: Warachate Khobjai, Suriyan Sukati, Khemjira Jarmkom, Pattaranut Eakwaropas, Surachai Techaoei

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The propose of this study was to investigate in vitro thrombolytic activity of Zingiber cassumunar Roxb. and Prasaplai, a Thai herbal formulation of Z. cassumunar Roxb. Herbs were extracted with boiling water and concentrated by lyophilization. To observe their thrombolytic potential, an in vitro clot lysis method was applied where streptokinase and sterile distilled water were used as positive and negative controls, respectively. Crude aqueous extracts from Z. cassumunar Roxb. and Prasaplai formula showed significant thrombolytic activity by clot lysis of 17.90% and 25.21%, respectively, compared to the negative control water (5.16%) while the standard streptokinase revealed 64.78% clot lysis. These findings suggest that Z. cassumunar Roxb. exhibits moderate thrombolytic activity and cloud play an important role in the thrombolytic properties of Prasaplai formula. However, further study should be done to observe in vivo clot dissolving potential and to isolate active component(s) of these extracts.

Keywords: thrombolytic activity, clot lysis, Zingiber cassumunar Roxb., Prasaplai formula, aqueous extract

Procedia PDF Downloads 298

Authors: Priyanka Yadav, Umesh Bnasal, Yashvinder Kumar

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The placenta is an important structure that provides oxygen and nutrients to the growing fetus in utero. It is usually thrown away after birth, but it has a therapeutic role in the regeneration of tissue. It is covered by the amniotic membrane, which can be easily separated into the amnion layer and the chorion layer—the amnion layer act as a biofilm for the healing of burn wound and non-healing ulcers. The freshly collected membrane has stem cells, cytokines, growth factors, and anti-inflammatory properties, which act as a biofilm for the healing of wounds. It functions as a barrier and prevents heat and water loss and also protects from bacterial contamination, thus supporting the healing process. The application of Amnion membranes has been successfully used for wound and reconstructive purposes for decades. It is a very cheap and easy process and has shown superior results to allograft and xenograft. However, there are very few case reports of amnion membrane grafting in newborns; we intend to highlight its therapeutic importance in burn injuries in newborns. We present a case of 9 days old male neonate who presented to the neonatal unit of Maulana Azad Medical College with a complaint of fluid-filled blisters and burns wound on the body for six days. He was born outside the hospital at 38 weeks of gestation to a 24-year-old primigravida mother by vaginal delivery. The presentation was cephalic and the amniotic fluid was clear. His birth weight was 2800 gm and APGAR scores were 7 and 8 at 1 and 5 minutes, respectively. His anthropometry was appropriate for gestational age. He developed respiratory distress after birth requiring oxygen support by nasal prongs for three days. On the day of life three, he developed blisters on his body, starting from than face then over the back and perineal region. At a presentation on the day of life nine, he had blisters and necrotic wound on the right side of the face, back, right shoulder and genitalia, affecting 60% of body surface area with full-thickness loss of skin. He was started on intravenous antibiotics and fluid therapy. Pus culture grew Pseudomonas aeuroginosa, for which culture-specific antibiotics were started. Plastic surgery reference was taken and regular wound dressing was done with antiseptics. He had a storming course during the hospital stay. On the day of life 35 when the baby was hemodynamically stable, amnion membrane grafting was done on the wound site; for the grafting, fresh amnion membrane was removed under sterile conditions from the placenta obtained by caesarean section. It was then transported to the plastic surgery unit in half an hour in a sterile fluid where the graft was applied over the infant’s wound. The amnion membrane grafting was done twice in two weeks for covering the whole wound area. After successful uptake of amnion membrane, skin from the thigh region was autografted over the whole wound area by Meek technique in a single setting. The uptake of autograft was excellent and most of the areas were healed. In some areas, there was patchy regeneration of skin so dressing was continued. The infant was discharged after three months of hospital stay and was later followed up in the plastic surgery unit of the hospital.

Keywords: amnion membrane grafting, autograft, meek technique, newborn, regeneration of skin

Procedia PDF Downloads 139

Authors: Deepanwita Deka, Dhruva Kumar Jha

Abstract:

Endophytes are the microbes that colonize living, internal tissues of plants without causing any immediate, overt negative effects. Endophytes are rich source of therapeutic substances like antimicrobial, anticancerous, herbicidal, insecticidal, immunomodulatory compounds. Brucea mollis, commonly known as Quinine in Assam, belonging to the family Simaroubaceae, is a shrub or small tree, recorded as endangered species in North East India by CAMP survey in 2003. It is traditionally being used as antimalarial and antimicrobial agent and has antiplasmodial, cytotoxic, anticancer, diuretic, cardiovascular effect etc. Being endangered and medicinal; this plant may host certain noble endophytes which need to be studied in depth. The aim of the present study was isolation and identification of potent endophytic fungi from Brucea mollis, an endangered medicinal plant, to protect it from extinction due to over use for medicinal purposes. Aseptically collected leaves, barks and roots samples of healthy plants were washed and cut into a total of 648 segments of about 2 cm long and 0.5 cm broad with sterile knife, comprising 216 segments each from leaves, barks and roots. These segments were surface sterilized using ethanol, mercuric chloride (HgCl2) and aqueous solution of sodium hypochlorite (NaClO). Different media viz., Czapeck-Dox-Agar (CDA, Himedia), Potato-Dextrose-Agar (PDA, Himedia), Malt Extract Agar (MEA, Himedia), Sabourad Dextrose Agar (SDA, Himedia), V8 juice agar, nutrient agar and water agar media and media amended with plant extracts were used separately for the isolation of the endophytic fungi. A total of 11 fungal species were recovered from leaf, bark and root tissues of B. mollis. The isolates were screened for antimicrobial, antioxidant and enzymatic activities using certain protocols. Cochliobolus geniculatus was identified as the most dominant species. The mycelia sterilia (creamy white) showing highest inhibitory activity against Candida albicans (MTCC 183) was induced to sporulate using modified PDA media. The isolate was identified as Geosmithia pallida. The internal transcribed spacer of rDNA was sequenced for confirmation of the taxonomic identity of the sterile mycelia (creamy white). The internal transcribed spacer r-DNA sequence was submitted to the NCBI (KU693285) for the first time from India. G. pallida and Penicillium showed highest antioxidant activity among all the isolates. The antioxidant activity of G. pallida and Penicillium didn’t show statistically significant difference (P˃0.05). G. pallida, Cochliobolus geniculatus and P. purpurogenum respectively showed highest cellulase, amylase and protease activities. Thus, endopytic fungal isolates may be used as potential natural resource of pharmaceutical importance. The endophytic fungi, Geosmithia pallida, may be used for synthesis of pharmaceutically important natural products and consequently can replace plants hitherto used for the same purpose. This study suggests that endophytes should be investigated more aggressively to better understand the endophyte biology of B. mollis.

Keywords: Antimicrobial activity, antioxidant activity, Brucea mollis, endophytic fungi, enzyme activity, Geosmithia pallida

Procedia PDF Downloads 150

Authors: Babak Rezavand

Abstract:

Pneumocystis carinii pneumonia (PCP) is one of the main causes of morbidity and mortality among immunocompromised and HIV-positive patients and remained one of the most important common opportunistic infections in these individuals in the world. Pneumocystis infection has been reported in many mammals. The aim of this study was to determine the Pneumocystis infection in wild rats as natural reservoirs of this organism in Tehran city, Iran. Fifty three rats (Rattus rattus) were live trapped in different areas of Tehran city, Iran. After isolation of their lung tissues and homogenization in sterile conditions, DNA was extracted. DNAs from all of the Pneumocystis species were amplified by pAZ102-H and pAZ102-E primers, and Nested PCR was performed using pAZ102-X and pAZ102-W primers from the initial PCR product for all the species of Pneumocystis. Amplification of the genome revealed the presence of Pneumocystis in the lungs of 17 rats (32%) through a PCR product with a bandwidth of 346 bp. In the Nested PCR amplification, from the PCR product of 53 rats, 64.2% of the samples were positive with a bandwidth of 261bp. Pneumocystis SPP infestation is highly prevalent among wild rats in Tehran city, indicating the existence of infection in the natural ecosystem of these rodents. As a host, rat plays an important role in the transmission of the microorganism in the world.

Keywords: pneumocystis SPP, rattus rattus, nested PCR, Tehran

Procedia PDF Downloads 178