Information about  S.aureus

Genetic Engineering

DNA Manipulating enzymes

 https://drive.google.com/open?id=1MWkdmE8MlVDtydH0QDpOVDBIATYyOADL&authuser=1

Agarose gel electrophoresis

https://www.researchgate.net/publication/224829869_Principles_of_Nucleic_Acid_Separation_by_Agarose_Gel_Electrophoresis

SDS PAGE

https://ruo.mbl.co.jp/bio/e/support/method/sds-page.html

http://howbiotech.com/the-principle-and-procedure-of-polyacrylamide-gel-electrophoresis-sds-page/

https://www.creativebiomart.net/blog/principle-and-protocol-of-sodium-dodecyl-sulphate-polyacrylamide-gel-electrophoresis-sds-page/

https://vlab.amrita.edu/?sub=3&brch=186&sim=319&cnt=1

https://capricorn.bc.edu/bi204/wp-content/uploads/2015/08/Chapter-14-2015.pdf

UV-Visible Spectrophotometry

Spectrofluorimetry

http://namrataheda.blogspot.com/2014/02/spectrophotometry-spectrofluorimetry.html
http://namrataheda.blogspot.com/2014/02/spectrophotometry-spectrofluorimetry_25.html

UV-Visible Spectrophotometry

http://namrataheda.blogspot.com/2013/07/spectrophotometry-part-2-uv-visible.html

HPLC

HPLC

Biological Model presentation 

By

II B.Sc. Microbiology Students 

in the 

National Symposium on Graduate Research (NSGR 2020)

Organized by 

Dr.  N.G.P. Arts and Science College 

Under 

DBT Star College Scheme 

on

21 and 22 Feb. 2020

 Group Photo with Models

 Explaining the working principle

DNA Model

 Corona Virus Model (COVID)

DNA, Bio-insecticide, Virus and Diffusion Model by II B.Sc. Micro students 

Ion Exchange Chromatography -study material

 Click below link

Ion Exchange Chromatography

Ion Exchange Chromatography video 

                                                       Click below link
                                                  Ion Exchange Chromatography

Industrial Visit Pasteur Institute

II B.Sc. Microbiology

Under 

DBT Star College Scheme  

 Lecture given about vaccine and uses by the Expert

Group photo in front of the Institution

Parasitism 

Parasitism

Workshop on Column Chromatography and Thin Layer Chromatography

Under DBT Star College Scheme on 01/02/2020

Organized by Department of Microbiology

Dr. N.G.P. Arts and Science College

Workshop Manual for Column and Thin Layer chromatography

Manual for column and Thin Layer Chromatography

Mushroom cultivation - A simple method 

Click below link

Mushroom cultivation method

Biofertilizer 

Click below link to get
the PPT

Biofertilizer

Treponema pallidum 

T.pallidum

Nitrogen fixation by Microorganisms

Air contains approximately 79% nitrogen, but plants cannot use it. Nitrogen is the most limiting nutrient for plant growth. The conversion of N2 into the compound which can be absorbed by the plants by biological means is termed as nitrogen fixation. Nitrogen is the major part of proteins and nucleic acids which are the basis of all life forms. It is also present in chlorophylls, alkaloids, and cytochromes. Nitrogen from the soil is continually depleted by soil erosion, chemical voltalisation, and denitrification etc. The bacteria present in the root nodules of the leguminous plant helps in restoring N2 in the soil. Significance of biological nitrogen fixation as the major mechanism of recycling of nitrogen from the unavailable atmospheric form to available forms in the biosphere is well documented.

Nitrogen fixation process is also influenced by soil temperature. The optimum temperature for nitrogen fixation is 55 to 80°C. N2 fixation takes place mainly in the specialized cell called heterocyst of the cyanobacteria but non-heterocystous cyanobacteria have also been reported to fix N2. The factors which affect this process is the level of soil nitrogen, the rhizobia strain to infect the legume, amount of legume plant growth and the length of growing season. The rate of nitrogen fixation depends upon the rate of plant growth. We may define N2 fixation as the conversion of atmospheric N2 into nitrogenous salt which is readily absorbed by the plant. Nitrogen present in the atmosphere in the dinitrogen (N=N) form, therefore, the microorganism which fixes nitrogen is called diazotrophs. Nitrogen fixation by the activity of microorganisms is called biological N2 fixation and the microorganisms are called biofertilizers. But N2 -fixation also occurs spontaneously due to lightening. The dinitrogen reduction process is mediated by nitrogenase. In the case of photosynthetic bacteria, nitrogen fixation with nitrogenase frequently involves hydrogen generation as a byproduct. Such hydrogen production by photosynthetic bacteria may be a viable source for fuel cells, a possible next-generation energy source that is, independent of fossil fuels (Wakayama et al., 2000).

Aerobic diazotrophs—require oxygen for growth and fix nitrogen in the presence of oxygen (low concentration) (Azotobacter, methane-producing bacteria).

Free-living diazotrophs—fix nitrogen both in aerobic and anaerobic conditions (Bacillus and Klebsiella).

Symbiotic diazotrophs—fix nitrogen only by the formation of nodules (Rhizobium, Bradyrhizobium and Sinorhizobium).

Biological nitrogen fixation can be explained in two ways i.e., symbiotic and asymbiotic N2 fixation.

Non-biological

Lightning heats the air around it breaking the bonds of N₂ starting the formation of nitrous acid. Nitrogen can be fixed by lightning that converts nitrogen and oxygen into NOx (nitrogen oxides). NOx may react with water to make nitrous acid or nitric acid, which seeps into the soil, where it makes nitrate, which is of use to plants. Nitrogen in the atmosphere is highly stable and nonreactive due to the triple bond between atoms in the N₂ molecule. Lightning produces enough energy and heat to break this bond allowing nitrogen atoms to react with oxygen, forming NOx. These compounds cannot be used by plants, but as this molecule cools, it reacts with oxygen to form NO₂ This molecule, in turn, reacts with water to produce HNO₃ (nitric acid), or its ion NO−3 (nitrate), which is usable by plants.

Biological

Biological nitrogen fixation was discovered by German agronomist Hermann Hellriegel and Dutch microbiologist Martinus Beijerinck. Biological nitrogen fixation (BNF) occurs when atmospheric nitrogen is converted to ammonia by a nitrogenase enzyme. The overall reaction for BNF is:

N₂+6e^-+12ATP+12H₂O                                 2NH4++12ADP+12Pi+4H⁺

                                        Nitrogenase complex 

The process is coupled to the hydrolysis of 16 equivalents of ATP and is accompanied by the co-formation of one equivalent of H₂. The conversion of N₂ into ammonia occurs at a metal cluster called FeMoco, an abbreviation for the iron-molybdenum cofactor. The mechanism proceeds via a series of protonation and reduction steps wherein the FeMoco active site hydrogenates the N₂ substrate. In free-living diazotrophs, nitrogenase-generated ammonia is assimilated into glutamate through the glutamine synthetase/glutamate synthase pathway. The microbial nif genes required for nitrogen fixation are widely distributed in diverse environments. Nitrogenases are rapidly degraded by oxygen. For this reason, many bacteria cease the production of the enzyme in the presence of oxygen. Many nitrogen-fixing organisms exist only in anaerobic conditions, respiring to draw down oxygen levels, or binding the oxygen with a protein such as leghemoglobin.

Microorganisms

Diazotrophs are widespread within domain Bacteria including cyanobacteria (e.g. the highly significant Trichodesmium and Cyanothece), as well as green sulfur bacteria, Azotobacteraceae, rhizobia and Frankia. Several obligately anaerobic bacteria fix nitrogen including many (but not all) Clostridium spp. Some archaea also fix nitrogen, including several methanogenic taxa, which are significant contributors to nitrogen fixation in oxygen-deficient soils.

Cyanobacteria inhabit nearly all illuminated environments on Earth and play key roles in the carbon and nitrogen cycle of the biosphere. In general, cyanobacteria can use various inorganic and organic sources of combined nitrogen, such as nitrate, nitrite, ammonium, urea, or some amino acids. Several cyanobacteria strains are also capable of diazotrophic growth, an ability that may have been present in their last common ancestor in the Archean eon. Nitrogen fixation by cyanobacteria in coral reefs can fix twice as much nitrogen as on land—around 660 kg/ha/year. The colonial marine cyanobacterium Trichodesmium is thought to fix nitrogen on such a scale that it accounts for almost half of the nitrogen fixation in marine systems globally.

Marine surface licing[check spelling] and non-photosynthetic bacteria belonging in Proteobacteria and Planctomycetes fixate significant atmospheric nitrogen.

Root nodule symbioses

Legume family

Plants that contribute to nitrogen fixation include those of the legume family—Fabaceae— with taxa such as kudzu, clover, soybean, alfalfa, lupin, peanut and rooibos. They contain symbiotic rhizobia bacteria within nodules in their root systems, producing nitrogen compounds that help the plant to grow and compete with other plants. When the plant dies, the fixed nitrogen is released, making it available to other plants; this helps to fertilize the soil. The great majority of legumes have this association, but a few genera (e.g., Styphnolobium) do not. In many traditional and organic farming practices, fields are rotated through various types of crops, which usually include one consisting mainly or entirely of clover or buckwheat (non-legume family Polygonaceae), often referred to as "green manure".

Non-leguminous

Other nitrogen fixing families include:

• Parasponia, a tropical genus in the family Cannabaceae, which are able to interact with rhizobia and form nitrogen-fixing nodules.
• Actinorhizal plants such as alder and bayberry can form nitrogen-fixing nodules, thanks to a symbiotic association with Frankia bacteria. These plants belong to 25 genera distributed across eight families.

The ability to fix nitrogen is present in other families that belong to the orders Cucurbitales, Fagales, and Rosales, which together with the Fabales form a clade of eurosids. The ability to fix nitrogen is not universally present in these families. For example, of 122 Rosaceae genera, only four fix nitrogen. Fabales were the first lineage to branch off this nitrogen-fixing clade; thus, the ability to fix nitrogen may be plesiomorphic and subsequently lost in most descendants of the original nitrogen-fixing plant; however, it may be that the basic genetic and physiological requirements were present in an incipient state in the most recent common ancestors of all these plants, but only evolved to full function in some of them.

Treponema pallidum

Treponema pallidum

Assignment on Vibrio cholarae 

Vibrio cholarae

Corynebacterium diptheriae

Corynebacterium diphtheriae

Students Poster presentation 

 "Power of Microorganisms"

 "Anton Van Leeuweenhok- Father of Microbiology"

 "DNA - Super life programmer"

Laboratory Diagnosis of Klebsiella 

Laboratory diagnosis of Klebsiella

Basic information about Bacillus 

 Bacillus

Cellulose decomposition

 Microbial decomposition of cellulose in soil

Cellulose decomposition:

·         Cellulose is relatively resistant polysaccharide, found in the cell wall of plant cell.

·         Cellulose is a linear polymer of β-D-glucose in which glucose units are linked together by β-1,4-glycosidic bond.

·         It is the most abundant organic matter found in nature. In plant it occurs in association with lignin and hemicellulose.

Mechanism of cellulose decomposition:

·         Pathway of cellulose decomposition follows series of enzymatic reactions.

·         Enzymes responsible for cellulose decomposition is cellulase.

·         Cellulase is a complex of three enzymes (ie. C1 enzyme, β-1,4-glucanase and β-1,4-glucosidase).

·         Series of enzymatic reaction occurs outside the microbial cell in which complex cellulose is decomposed into free glucose molecules by extracellular enzymes.

Step I: hydrolysis by C1 enzymes:

C1 enzyme hydrolyses native cellulose polymer to form smaller fragments.

C1 enzyme is only found in true cellulolytic microorganisms.

Step II: hydrolysis by β-1,4-glucanase enzyme:

β-1,4-glucanase hydrolyze the smaller fragments of cellulose to form even smaller fragments such as disaccharides, tri-saccharides etc.

There are two types of glucanase ie. Endo-glucanase and Exo-glucanase.

Endo-glucanase randomly cuts the fragments somewhere in the middle whereas exo-glucanase sequentially release glucose molecule from one end of the fragment.

Some free glucose unit as well as disaccharides, tri-saccharides and other oligosaccharides are produced by the action of β-1,4-glucanase.

Step III: hydrolysis by β-1,4-glucosidase enzyme:

β-1,4-glucosidase hydrolyses di, tri and oligosaccharides to form free glucose molecules.

Step IV: metabolism of glucose:

Free glucose molecules then enter into microbial cell and metabolized by glycolysis to form pyruvate.

Depending upon types of microorganisms and the condition of environment, pyruvate is converted into CO2 and water or ethanol or any organic acids

Examples of Cellulolytic microorganisms; Cellulose decomposers

Bacteria: Bacillus, Cellulomonas, Clostridium, Cytophaga, Polyangium, Pseudomonas etc

Fungi: Aspergillus, Alterneria, Fomes, Fusarium, Myrothecium etc

Actinomycetes: Micromonospora, Nocardia, Streptomyces, Streptosporangium etc.

Factors affecting cellulose decomposition in soil:

Various environmental and other factors affects rate of cellulose decomposition in soil by microorgansims, some of them are;

i. Addition of available Nitrogen:

Additionof inorganic nitrogen compounds such as ammonia, nitrite or easily decomposable nitrogen compounds like aminoacids and proteins increase the rate of cellulose decomposition by microorganisms.

Microorganisms require both carbon and nitrogen for biosynthesis of their cellular materials. Therefore, microbial decomposition of cellulose cannot occurs without nitrogenous sources.

ii. Temperature:

Cellulose decomposition can occurs from temperature near freezing to above 65°C because both psychrophiles and thermophiles are involved in cellulose degradation.

But rate of cellulose decomposition is maximum in mesophilic range of temperature of 25-30°C because most cellulolytic microbes are mesophiles.

iii. Aeration:

In anaerobic soil, anaerobic bacteria like Clostridium decompose cellulose and in aerobic soli mainly fungi and aerobic bacteria take part in decomposition of cellulose.

Rate of cellulose is higher in aerobic soil.

iv. Moisture:

Excessive moisture brings anaerobic condition in soil. Therefore, rate of cellulose decomposition is slower in water logged soil.

v. pH:

In neutral to alkaline soil, bacteria and actinomycetes mainly take part in cellulose decomposition.

In acidic soil, fungi are dominant cellulose decomposers.

Rate of cellulose decomposition is slightly higher in acidic soil than alkaline and neutral.

vi. Addition of organic matter:

Addition of easily decomposable organic matters increase the rate of cellulose decomposition.

If only cellulose is present in soil, microorganisms cannot multiply fast so that the rate of decomposition become slower.

If small amount of easily decomposable organic matter is added initially in soil, microorganisms rapidly multiplies and grow their numbers. Furthermore cellulose is degraded rapidly when the easily decomposable organic matter is exhausted.

vii. Lignin:

Lignin slows the rate of cellulose decomposition.

Lignin itself is not toxic to cellulolytic microbes. Inhibition of cellulose decomposition by lignin is due to its close association to cellulose in cell wall.

Control of Epidemis 

Control of epidermis

Klebsiella - Brief information 

Klebsiella - Brief information