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Isolation and enrichment of bacteriophages by membrane filtration immobilization technique.(Current protocol in cell Biology)
(Ghugare, G. S.,Nimkande, V
,e41. ,,,Year : 2017)
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The method described here enables rapid bacteriophage isolation and enrichment of host-specific bacteriophages from an environmental sample. This is achieved by using a simple 0.45-µm Milli pore membrane where a specific host is immobilized on the membrane and a sample suspected of containing bacteriophages is exposed to the immobilized cells with the help of a membrane ltration unit. This filtration step facilitates host-specific interaction of bacteriophages with the host and maximization of this interaction using a classic membrane filtration method.Under the effect of vacuum from a vacuum pump, a filter assembly provides a chance for every bacteriophage in the sample to interact with the specific host on the membrane filter. Our technique allows retaining specific bacteriophages on the membrane along with its host cells via adsorption; these adsorbed bacteriophages (along with their hosts) on a filter disc are then enriched in regular nutritive broth, tryptone soya broth (TSB), by incubation. With help of a plaque assay method, host-specific phages of various bacterial species can be isolated, segregated, and enriched
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Isolation of bacteriophages from air using vacuum filtration technique: an improved and novel method.(Journal of applied microbiology)
(Magare B, Nair A,
,Volume 123,, Issue 4 ,Page 896–902,Year : 2017)
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"AIM: Development of a simple and economical air sampler for isolation and enrichment of bacteriophages from air samples. METHODS AND RESULTS: A vacuum filtration unit with simple modifications was used for isolation of bacteriophages from air sampled in the lavatory. Air was sampled at the rate of 62 l min-1 by bubbling into Mcllvaine buffer for 30 min, which was used as bacteriophage solution for enrichment and plaque assessment against individual hosts. Alternatively, the aforementioned phage solution was enriched using a host consortium before plaque assessment. Phages were isolated in the range of 1-12 PFU per ml by the first method, whereas enrichment with host consortium gave phages around 10- to 1000-folds higher in number. Combining with established enrichment method, an improvement of about 10 times in phage isolation efficiency was attained. CONCLUSIONS: The method is very useful for studying the natural bacteriophages of air, requiring only a basic microbiological laboratory setup making it simple and economical. SIGNIFICANCE AND IMPACT OF THE STUDY: This study brings out a simple, economical air sampler for assessing air bacteriophages that can be employed by any microbial laboratory. Although various methods are available for studying bacteriophages in water and soil, very limited are available for air. To the best of our knowledge, the method developed in this study is unique in its design and concept for studying bacteriophages in air. The sampler is sterilizable by autoclaving and maintains a healthy rate of airflow provided by conventional vacuum pumps. The use of a nonspecific 'trapping solution' allows for the qualitative and quantitative study of air bacteriophages."
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Dielectric Barrier Discharge based Mercury-free plasma UV-lamp for efficient water disinfection. (Nature Scientific Reports)
(Ram Prakash, Afaque M. Hossain, U.N. Pal and M. Krishnamohan
,,,,Year : 2017)
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No information is available
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Ganges: special at its origin.(Journal of Biological Research-Thessaloniki)
(Khairnar, Krishna,,,16,Year : 2016)
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No information is available
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Membrane filtration immobilization techniquea simple and novel method for primary isolation and enrichment of bacteriophages(Journal of applied microbiology)
(Ghugare, G. S., Nair, A., Nimkande, V., Sarode, P., Rangari, P.
,Vol.122 ,Issue 2,531-539,Year : 2016)
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Aim:
To develop a method for the isolation and enrichment of bacteriophages selectively against speciÞc bacteria coupled with a membrane Þltration technique. Methods and Results: Rapid isolation and concentration of host-speciÞc bacteriophages was achieved by exposure of the sample suspected to contain bacteriophages to a speciÞc host immobilized on a 0 45 lm membrane in a membrane Þltration unit. The principle behind this method is the exploitation of host-speciÞc interaction of bacteriophages with their host and maximizing this interaction using a classic membrane Þltration method. This provides a chance for each bacteriophage in the sample to interact with the speciÞc host on the membrane Þlter Þtted with a vacuum pump. SpeciÞc bacteriophages of the host are retained on the membrane along with its host cells due to the effect of adsorption and these adsorbed bacteriophages (along with their hosts) on the Þlter disc are then ampliÞed and enriched in regular nutritive broth tryptose soya broth by incubation. With the help of the plaque assay method, host-speciÞc phages of various bacterial species were isolated, segregated and enriched.
Conclusions:
The phage concentration method coupled with membrane Þltration immobilization of host bacteria was able to isolate and enrich the host-speciÞc bacteriophages by several fold using a lower quantity of an environmental water sample, or other phage suspensions. Enrichment of phages from single plaques was also achieved. Significance and Impact of the Study: The isolation and detection of host-speciÞc bacteriophages from a low density bacteriophage water sample in a single step by the use of a simple and basic microbiological technique can be achieved. Enrichment of phages from low phage titre suspensions is also achieved very effectively.
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Novel application of bacteriophage for controlling foaming in wastewater treatment plant-an eco-friendly approach(Bioengineered)
(Pal P, Chandekar RH
,7(1),,46-49,Year : 2016)
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This addendum to "Novel application of bacteriophage for controlling foaming in wastewater treatment plant- an eco-friendly approach " includes characteristics of the phages NOC1, NOC2 and NOC3 not discussed in the previous paper. The phage adsorption and host interaction properties, their sensitivity to pH and temperature are inferred. NOC2 is seen to be more temperature resistant while others are not. All the phages show pH sensitivity. There is a variance observed in the behavior of these phages. Also, applicability of the phage based system to large scale reactors is studied and discussed here.
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Re-emergence of Swine flu in Central India, 2014. (Indian Pediatrics)
(Dhande L
, 52(1),,77,Year : 2015)
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No information is available
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Causes and Remedies For Filamentous Foaming In Activated Sludge Treatment Plant.(Global NEST Journal)
(Khairnar K, Pal P, Paunikar W,Vol 16,,,p 762-772,Year : 2014)
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"This paper reviews the problem of foaming associated with the activated sludge process and its control using various physical, chemical and biological methods. Activated sludge process is widely used for treatment of every type of wastewater like industrial, domestic and municipal wastewater. This process is driven by a complex microbial population, among which some mycolic acid containing bacteria leads to the stable foam formation which ultimately results in poor efficiency of the plants and leading to major environmental, operational, and health problems. A number of researches provide the evidences of foaming in wastewater treatment plants and its control using physical, chemical and biological methods. Current approaches for controlling foam includes operational adjustments, additional structures, controlling dissolved oxygen levels, water sprays, steam application, polymer addition, chlorination and a novel and ecofriendly approach that is treatment of filamentous bacteria with the specific phages. A detailed study of all methods is presented and collectively described in this review paper for a better understanding of the foam controlling strategies. "
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A simple and novel modification of Comet assay for determination of bacteriophage mediated bacterial cell lysis.(Journal of virological methods)
("Khairnar K, Sanmukh S,Chandekar R,PaunikarW ,Volume 203,,, Page 33–38.,Year : 2014)
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The comet assay is the widely used method for in vitro toxicity testing which is also an alternative to the use of animal models for in vivo testing. Since, its inception in 1984 by Ostling and Johansson, it is being modified frequently for a wide range of application. In spite of its wide applicability, unfortunately there is no report of its application in bacteriophages research. In this study, a novel application of comet assay for the detection of bacteriophage mediated bacterial cell lysis was described. The conventional methods in bacteriophage research for studying bacterial lysis by bacteriophages are plaque assay method. It is time consuming, laborious and costly. The lytic activity of bacteriophage devours the bacterial cell which results in the release of bacterial genomic material that gets detected by ethidium bromide staining method by the comet assay protocol. The objective of this study was to compare efficacy of comet assay with different assay used to study phage mediated bacterial lysis. The assay was performed on culture isolates (N=80 studies), modified comet assay appear to have relatively higher sensitivity and specificity than other assay. The results of the study showed that the application of comet assay can be an economical, time saving and less laborious alternative to conventional plaque assay for the detection of bacteriophage mediated bacterial cell lysis.
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Isolation and characterization of bacteriophages infecting nocardioforms in wastewater treatment plant. (Biotechnology Research International)
(Khairnar K*, Pal P, Chandekar RH, Paunikar WN.
,151952.,,,Year : 2014)
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Activated sludge plants (ASP) are associated with the stable foaming problem worldwide. Apart from the physical and chemical treatment methods, biological treatment method has been least explored and may prove to be a novel and ecofriendly approach to tackle the problem of stable foam formation. In ASP Nocardia species are commonly found and are one of the major causes for forming sticky and stable foam. This study describes the isolation and characterization of three Nocardia bacteriophages NOC1, NOC2, and NOC3 for the control of Nocardia species. The bacteriophages isolated in this study have shown promising results in controlling foam producing bacterial growth under laboratory conditions, suggesting that it may prove useful in the field as an alternative biocontrol agent to reduce the foaming problem. To the best of our knowledge to date no work has been published from India related to biological approach for the control of foaming.
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Causes and Remedies For Filamentous Foaming In Activated Sludge Treatment Plant.(Global NEST Journal)
(Pal P
,Vol 16, 4,762-772,Year : 2014)
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No information is available
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