5.1. INTRODUCTION Contamination by …shodhganga.inflibnet.ac.in/bitstream/10603/39782/11/11...ropiness a sticky, stringy consistency caused by bacterial production of long-chain polysaccharides

Chapter 5 Antimicrobial Study

Dept. of Chemistry, SPU 191

5.1. INTRODUCTION

Contamination by microorganisms is of great concern in areas of medical

devices, healthcare products, water purification systems, hospital and

dental office equipments, food packaging and food storage industries etc.

One possible way to avoid the microbial contamination is to develop

materials possessing antimicrobial activities. Consequently, biocidal

polymers have received much attention in recent years [1]. The

antimicrobial polymers are suitable in variety of applications such as film

and packaging material, foodstuffs, containers, in sanitary applications

and many other consumer’s including common consumer’s applications

like cosmetic, medical equipment and other devices.

The synthesis of macrocyclic ligands and their metal complexes is

a growing area of research in inorganic and bioinorganic chemistry. The

synthesis and study of macrocycles have undergone tremendous growth

in recent years and their complexation chemistry with a wide variety of

metal ions has been extensively studied. Macrocycles find wide

applications in medicine, cancer diagnosis, in treatment of tumors and

treatment of kidney stone.

The use of simple inorganic complexes is the example of applied

bioinorganic chemistry. There are areas of chemistry which are not

primarily biologically oriented, however, such area make use of the

bioinorganic chemistry. Now a days, the biochemists have begun to

investigate the molecular details of enzymes and biologically active

compounds, particularly metal complexes. Coordination compounds play

an important role in various biochemical processes. The inorganic

chemists on the other hand have slowly begun to recognize the similarity

between the compounds they synthesized and biologically important

compounds containing metal ions. This is an active area of research of

bioinorganic chemistry [2]. This discipline is rapidly bridging the gap

between traditional inorganic chemistry and biochemistry. Sigel and

McCormick have discussed the discriminating behaviour of metal ions

and ligands with respect to their biological significance [3]. They

attempted to answer the question: Which are the control mechanisms that

determine the coordination and coordination tendency of the metal ions?

During the past four decades, considerable progresses in understanding



the role of metals in biological process has resulted from the discovery of

a large number of metalloproteins and performs experiments to ascertain

their function [4].

The study of the antimicrobial activity of lanthanides and their

coordination polymers has great significance due to their biological and

clinical aspects [5]. The biological properties of the lanthanides based on

their similarity to calcium, have stimulated research into their

therapeutic applications.

The polychelates of phenols possess interesting and excellent

microbial activities like growth inhibition. Polymeric coordinating

reagents are a novel type of substances possessing a combination of

physical properties of a polymer and chemical properties of the attached

reagent. When organic polymers are used as adhesive coatings, some of

them can be infected by microorganisms such as bacteria and fungi [6].

This problem can be solved by the addition of metal ions in the polymeric

system. This changes the physicochemical properties of the polymer and

restricts the attack of microorganism. Therefore, coordination polymers

are also used as biocidal coatings and also widely applied to prevent the

growth of microorganism on surfaces. e. g. antifouling paints [7].

In the present study, bacterial strains such as Escherichia coli,

Bacillus subtilis and Staphylococcus aureus, yeast strains Saccharomyces

cerevisiae were used as test bio-organism for the biocidal activity testing.

Accordingly their general information regarding their history and living

style are briefly discussed.

5.2. BACTERIA AND YEAST

5.2.1. Bacteria

In 1838, a German scientist Ehrenberg C. E. used the term “bacterium”.

Bacteria are the microscopic organisms of plant kingdom and are devoid

of chlorophyll. They are relatively simple and primitive form of cellular

organisms known as “Prokaryotes”. Bacteriology is the science that deals

with the study of bacteria.

The Danish Physician Christian Gram in 1884, discovered a stain

known as Gram stain, which can divide all bacteria into two classes



“Gram positive” and “Gram negative”. The Gram-positive bacteria resist

decolouration with acetone, alcohol and remain stained (methyl violet) as

dark blue coloured, while Gram-negative bacteria are decolorized.

Bacteria can be classified according to their morphological characteristics

as lower and higher bacteria. The lower bacteria have generally

unicellular structures, never in the form of mycelium or sheathed

filaments, e.g., cocci, bacilli, etc. The higher bacteria are filamentous

organisms, few being sheathed having certain cells specialized for

producing diseases in animal or human, are known as “Pathogens”.

The organisms were identified by using the following stains.

Schiff technique periodic acid

Gram stains

Zeil Nelsonm acid fast stains

Classification:

Escherichia Coli is species of schizomycetes class; having Eubacterial

order, Enterobacteriaceae family and Escherichia genus.

Bacillus Subtillis is species of schizomycetes class; having

Eubacterials order, Bacteriodaceac family and fusobacterium

streptobacillus and sphaerophorus genus.

Staphylococcus Aureus is species of schizomycetes class; having

Eubacterials order, micrococeaceac family and staphylococcus genus.

5.2.1.1. Escherichia Coli

Escherichia coli is a gram negative bacterium that is commonly found in

the lower intestine of warm-blooded animals. Most E. coli strains are

harmless, but some, such as serotype O157:H7, can cause serious food

poisoning in humans and are occasionally responsible for costly product

recalls [8, 9]. The harmless strains are part of the normal flora of the gut,

and can benefit their hosts by producing vitamin K2 [10] or by preventing

the establishment of pathogenic bacteria within the intestine [11, 12].

E. coli are not always confined to the intestine and their ability to

survive for brief periods outside the body makes them an ideal indicator

organism to test environmental samples for fecal contamination [13, 14]



The bacteria can also be grown easily and its genetics are comparatively

simple and easily-manipulated, making it one of the best-studied

prokaryotic model organisms and an important species in biotechnology.

E. coli was discovered by German pediatrician and bacteriologist Theodor

Escherich in 1885 [13] and is now classified as part of the

Enterobacteriaceae family of gamma-proteobacteria [15]

5.2.1.2. Bacillus Subtilis

Bacillus subtilis is a Gram-positive, catalase-positive bacterium commonly

found in soil [16]. A member of the genus Bacillus, B. subtilis is rod-

shaped, and has the ability to form a tough, protective endospore,

allowing the organism to tolerate extreme environmental conditions.

Unlike several other well-known species, B. subtilis has historically been

classified as an obligate aerobe, though recent research has demonstrated

that this is not strictly correct [17]. B. subtilis has also been called the

Hay or Grass bacillus; Bacilluss globigii or Bacillis licheniformis are

binomial synonyms [18, 19].

B. subtilis is not considered a human pathogen; it may contaminate

food but rarely causes food poisoning [20]. B. subtilis produces the

proteolytic enzyme subtilisin. B. subtilis spores can survive the extreme

heating that is often used to cook food, and it is responsible for causing

ropiness — a sticky, stringy consistency caused by bacterial production of

long-chain polysaccharides — in spoiled bread dough.

5.2.1.3. Staphylococcus Aureus

S. aureus is a Gram-positive coccus, which appears as grape-like clusters

when viewed through a microscope and has large, round, golden-yellow

colonies, often with β-hemolysis, when grown on blood agar plates [21].

Clinically, the most important genus of the Micrococcaceae family is

Staphylococcus. The Staphylococcus genus is classified into two major

groups: aureus and non-aureus. S. aureus is a leading cause of soft tissue

infections, as well as toxic shock syndrome (TSS) and scalded skin

syndrome. It can be distinguished from other species of Staphylococcus

by a positive result in a coagulase test (all other species are negative).



The pathogenic effects of Staphylococcus are mainly associated

with the toxins it produces. Most of these toxins are produced in the

stationary phase of the bacterial growth curve. In fact, it is common for

an infected site to contain no viable Staphylococcus cells. The S. aureus

enterotoxin causes quick onset food poisoning which can lead to cramps

and severe vomiting. Infection can be traced to contaminated meats

which have not been fully cooked. These microbes also secrete

leukocidin, a toxin which destroys white blood cells and leads to the

formation of pus and acne. Particularly, S. aureus has been found to be

the causative agent in such ailments as pneumonia, meningitis, boils,

arthritis and osteomyelitis (chronic bone infection).

5.2.2. Yeast

Yeasts are unicellular microorganisms classified in the kingdom Fungi,

with about 1,500 species currently described [22]; they dominate fungal

diversity in the oceans.

The word "YEAST" comes from Old English gist, gyst, and from the

Indo-European root yes-, meaning boil, foam, or bubble. Yeast microbes

are probably one of the earliest domesticated organisms. People have

used yeast for fermentation and baking throughout history. In 1680 the

Dutch naturalist Antonie van Leeuwenhoek first microscopically observed

yeast, but at the time did not consider them to be living organisms but

rather globular structures. In 1857 French microbiologist Louis Pasteur

proved in the paper "Mémoire sur la fermentation alcoolique" that

alcoholic fermentation was conducted by living yeasts and not by a

chemical catalyst.

Yeasts are characterized by a wide dispersion of natural habitats.

Common on plant leaves and flowers, soil and salt water. Yeasts are also

found on the skin surfaces and in the intestinal tracts of warm-blooded

animals, where they may live symbiotically or as parasites.

5.2.2.1. Saccharomyces Cerevisiae

Saccharomyces cerevisiae is a species of budding yeast. It is perhaps the

most useful yeast owing to its use since ancient times in baking and



brewing. It is believed that it was originally isolated from the skins of

grapes (one can see the yeast as a component of the thin white film on

the skins of some dark-colored fruits such as plums; it exists among the

waxes of the cuticle). It is one of the most intensively studied eukaryotic

model organisms in molecular and cell biology, much like Escherichia coli

as the model prokaryote. It is the microorganism behind the most

common type of fermentation. Saccharomyces cerevisiae cells are round

to ovoid, 5-10 micrometers in diameter. It reproduces by a division

process known as budding.

5.3. CHARACTERISTICS OF USEFUL ANTIMICROBIAL AGENTS

There is not a single chemical agent which is best for the control of

microorganisms for any and all the purposes. A general purpose chemical

antimicrobial agent would have an extremely elaborate array of

characteristics [23] as mentioned below:

Broad spectrum of antimicrobial activity at lowest concentration.

Solubility, Stability, Homogeneity, Availability.

Non combination with extraneous organic materials, capacity to

penetrate.

Toxic to microorganisms at room or body temperature.

Deodorizing ability, detergent capacities.

5.4. EVALUATION OR SCREENING OF ANTIMICROBIAL AGENTS

5.4.1. Conditions Necessary for Inhibition or Control of

Microorganisms

The following conditions must be met for the screening of antimicrobial

activity:

There should be an intimate contact between test organisms and

substance to be evaluated.

Required conditions should be provided for the growth of

microorganisms.

Conditions should be same throughout the study.

Aseptic/sterile environment should be maintained.



5.4.2. Environmental or Physical Conditions

The physical or chemical properties of the medium carrying the

microorganism (i.e. environment) has a profound influence on the rate as

well as on the efficacy of microbial destruction, e.g. the temperature

influence is much greater in acid material as well as increase in

temperature may cause destruction, viscous nature of material will

markedly influence the penetration of the agent, presence of extraneous

organic matter can significantly reduce the efficacy of antimicrobial

agents by inactivating it or protecting microorganisms from agent.

5.4.3. Factors Affecting Activity of Antimicrobial Agents

Microorganisms are not simple physical targets. Many biological

characteristics influence the mode of action by various agents and factors

must be considered in the application of chemical agent used to inhibit or

destroy microbial populations. The main factors which influence the

efficiency of Antimicrobial agents are:

Nature of the chemotherapeutic agent

Types of microorganisms

Environmental conditions- such as chemical and physical properties of

medium or substance carrying the organisms, presence of extraneous

matter and temperature control.

5.4.4. Factors Affecting Inhibition Zone

5.4.4.1. Ingredient of Culture Media

Many substances are present in culture media, which may affect the zone

of inhibition. Common ingredients such as peptone, agar, etc. may vary in

their contents and many of these minerals may influence the activity of

some antimicrobials. It is well known that Ca, Mg, Fe, etc. ions affect the

sensitivity of zone produced by the tetracycline, gentamycin, NaCl reduce

the activity of amino glycosides and enhances the effect of fucidine.



5.4.4.2. Choice of Media

Consistent and reproducible results are obtained in media prepared

especially for sensitivity testing; the plates must be poured flat with an

even depth.

5.4.4.3. Effect of pH

The activity of amino glycosides is enhanced in alkaline media and

reduced in acidic media, the reverses is shown by tetracycline.

5.4.4.4. Size of Inoculums

Although large numbers of organisms do not markedly affect many

antibiotics, all inhibition zones are diminished by heavy inocula. The

ideal inoculum is one, which gives an even dense growth without being

confluent. Overnight broth cultures of organisms and suitable

suspensions from solid media can be diluted accurately to give optimum

inocula for sensitivity testing.

5.5. ACTION OF ANTIMICROBIAL AGENTS ON MICROORGANISMS

The manner in which antimicrobial agents inhibit or kill the microbial

growth can be attributed to the following kinds of action:

Inhibition of cell wall or damage to the cell wall.

Damage to the cytoplasmic membrane. - Alteration in the

permeability of the cytoplasmic membrane.

Inhibition of nucleic acid and protein synthesis.

Change in the physical and chemical state of proteins and nucleic

acids.

Inhibition of specific enzyme action.

5.6. EXPERIMENTAL

Various laboratory methods have been used from time to time by several

researchers for evaluation of antimicrobial activity. They are as follows:



1. Turbidometric method,

2. Agar streak dilution method,

3. Serial dilution method and

4. Agar diffusion method.

Agar diffusion method is again of three types:

i. Agar cup method,

ii. Agar ditch method and

iii. Paper disc Method.

In the present study, Agar Diffusion Method has been used to determine

antimicrobial activity.

5.6.1. Microorganisms

The bacterial strains of Escherichia coli, Bacillus substilis, Staphylococcus

aureus and yeast strain of Snccharomyces cerevisiae were tested with

polymeric ligands and their polychelates with Ln(III). The effect of the

compound in the growth media were investigated by standard

microbiological parameters. The bacterial culture was maintained on N-

agar (N-broth, 2.5% w/v agar). The yeast culture was maintained on MGYP

in 3% (w/v) agar agar, malt extract 0.3% (w/v), glucose 1.0% (w/v), yeast

extract 0.3% (w/v) and peptone 0.5% (w/v) on distilled water and the pH

was adjusted to 6.7-7.3. All were subcultured every fortnight and stored

at 0-5OC.

5.6.2. Composition of the Medium for Bacteria and Yeast

Nutrient Agar Medium Nutrient Broth Medium

Composition: Composition:

Beef Extract : 1.5gm Beef Extract : 2.5gm

Peptone : 6.0gm Peptone : 2.5gm

NaCl : 1.5gm NaCl : 1.25gm

Agar : 20gm Agar : 250gm

Distilled water : 1000mL Distilled water : 250mL

pH adjusted to : 6.7-7.3 pH adjusted to : 7.5

Both the above mentioned media were used and found to be suitable for

the growth of both organisms used in present work.



Composition of the medium for yeast

Yeast extract : 3gm

Malt extract : 3gm

Peptone : 5gm

Glucose : 10gm

Agar : 20gm

Distilled water : 1000mL

pH adjusted to : 5.5

5.6.3. Slant Preparation

Nutrient agar media dissolved in distilled water and was sterilized by

autoclave. About 5mL of molten media was transferred in previously

sterilized test tubes. The test tubes were then plugged tightly and were

placed in a slanting position to cool and solidify.

5.6.4. Stock Culture

Culture was grown on nutrient agar slants by incubating for 24 hrs at

37OC.

5.6.5. Inoculum Preparation of Bacteria and Yeast

Bacterial and Yeast culture, a loop of cell mass from pregrown slants was

inoculated into sterile N-broth tubes containing 15mL medium and

incubated on a shaker at 150 rpm and 37OC for 24 hrs to obtain sufficient

cell density (i.e. 1 X 108 cells/mL).

5.6.6. Preparation of the Solution

Antibacterial activity is usually tested by making aqueous solution of the

compounds. However, polychelates used in the present study were

insoluble in water, but soluble in dimethyl sulphoxide. Therefore, to

study antibacterial activity of the polychelates, their solutions were

prepared by using DMSO (dimethyl sulphoxide), may have no

antimicrobial activity, therefore, blank experiment was carried out with

DMSO and tested.



5.6.7. Screening of Compounds for their Antimicrobial Activity

Antimicrobial activity was checked by the Agar Diffusion Method [24].

Sterile, melted N-agar was poured into a sterile empty petri plate and

allowed to solidify. A ditch was prepared with the help of a sterile scalpel

on opposite ends, one for control (solvent without compound) and the

other for the test sample. The pregrown cultures were streaked parallel

from one ditch to another. One of the ditches was filled with respective

components dissolved in DMSO at concentrations ranging from 50-1000

ppm. Then after, the plates were transferred to the refrigerator for 10

minutes to allow the sample diffuse out from the ditch and into the agar

before organisms start growing followed by incubation at 37OC for 24 hrs.

Next day the distance in millimeter (mm), from the ditch was measured as

a parameter of inhibition. The polymeric ligands (resins) and their

polychelates were studied for their antimicrobial activity against standard

bacterial strains of E. coli, B. substillis, S. aureus (bacteria) and S.

cerevisiae (yeast). The compounds were tested at different concentrations

ranging from 50-1000 ppm to find out the minimum inhibitory

concentration (MIC) of the polymeric ligands (resins) and polychelates,

which inhibits the microbial growth [25]. The minimum concentration 500

ppm was found. The inhibition of growth from the ditch was measured in

millimeter (mm) and the results are shown in Tables 5.1 to 5.5.

5.7. RESULTS AND DISCUSSION

The polymeric ligand and their polychelates were studied for their

antimicrobial activity against standard bacterial strains of Escherichia

coli, Bacillus subtilis, Staphylococcus aureus (bacteria) and Saccharomyces

cerevisiae (yeast). The minimum concentration 500 ppm was found. The

inhibition of growth from the ditch was measured in millimeter (mm) and

the results are shown in Table 5.1 to 5.5. The polymeric ligand was found

biologically active and their polychelates showed significantly enhanced

antibacterial activity against one or more bacterial species, in comparison

to the uncomplexed polymeric ligand. It is known that chelation tends to

make the ligands act as more potent bactericidal agents, than the parent

ligand. The antimicrobial activity of the compounds increases after



chelation. Chelation reduces the polarity of the central metal ion by

partial sharing of its positive charge with the donor groups [26],

increasing lipophilic nature of the central metal ion, which in turn favors

its permeation to the lipid layer of the membrane. Other factors, viz.,

stability constant, molar conductivity, solubility and magnetic moment,

are also responsible for increase in the anti-microbial activity of the

polychelates [27].



5.7.1. Antimicrobial Activity of HEAP-EG Resin and its Polychelates

For the inhibition of E.coli (gram negative bacteria), B. substilis (gram

positive bacteria) and S.aureus (gram positive bacteria), HEAP-EG resin

was least active while, for S. cerevisiae (yeast), it was (10 mm) weakly

active.

For the inhibition of E.coli (gram negative bacteria), [La(III)-(HEAP-EG)] (16

mm), [Pr(III)-(HEAP-EG)] (17 mm), [Nd(III)-(HEAP-EG)] (16 mm), [Sm(III)-

(HEAP-EG)] (18 mm), [Tb(III)-(HEAP-EG)] (16 mm ) and [Dy(III)-(HEAP-EG)]

(18 mm) were significantly active, while [Gd(III)-(HEAP-EG)] (15 mm) was

moderately active.

For the inhibition of B. substilis (gram positive bacteria), [La(III)-(HEAP-EG)]

(20 mm), [Pr(III)-(HEAP-EG)] (22 mm), [Nd(III)-(HEAP-EG)] (18 mm), [Sm(III)-

(HEAP-EG)] (19 mm), [Gd(III)-(HEAP-EG)] (17 mm ), [Tb(III)-(HEAP-EG)] (22

mm) and [Dy(III)-(HEAP-EG)](21 mm) were significantly active.

For the inhibition of S.aureus (gram positive bacteria), [La(III)-(HEAP-EG)]

(19 mm), [Pr(III)-(HEAP-EG)] (18 mm), [Nd(III)-(HEAP-EG)] (22 mm), [Sm(III)-

(HEAP-EG)] (20 mm), [Gd(III)-(HEAP-EG)] (21 mm ), [Tb(III)-(HEAP-EG)] (19

mm) and [Dy(III)-(HEAP-EG)](20 mm) were significantly active.

For the inhibition of S. cerevisiae (yeast), [La(III)-(HEAP-EG)] (22 mm),

[Pr(III)-(HEAP-EG)] (23 mm), [Nd(III)-(HEAP-EG)] (20 mm), [Sm(III)-(HEAP-EG)]

(19 mm), [Gd(III)-(HEAP-EG)] (22 mm ), [Tb(III)-(HEAP-EG)] (20 mm) and

[Dy(III)-(HEAP-EG)](22 mm) were significantly active.



Table 5.1. Antimicrobial Activity data of the Polymeric Ligand (HEAP-EG)

and its Polychelates

Zone of inhibitiona (mm)

Ligand / Polychelates E.

coli

B.

substilis

S.

aureus

S.

cerevisiae

(HEAP-EG)n

-- -- -- 10

{[La(HEAP-EG)2(H

2O)

2}.OH]

n 16 20 19 22

{[Pr(HEAP-EG)2(H

2O)

2}.OH]

n 17 22 18 23

{[Nd(HEAP-EG)2(H

2O)

2}.OH]

n 16 18 22 20

{[Sm(HEAP-EG)2(H

2O)

2}.OH]

n 18 19 20 19

{[Gd(HEAP-EG)2(H

2O)

2}.OH]

n 15 17 21 22

{[Tb(HEAP-EG)2(H

2O)

2}.OH]

n 16 22 19 20

{[Dy(HEAP-EG)2(H

2O)

2}.OH]

n 14 21 20 22

DMSOb -- -- -- --

a16 – 23 mm = significantly active; 10 – 15 mm = moderately active;

< 10 mm = weakly active. bsolvent (negative control).



5.7.2. Antimicrobial Activity of HEAP-1,2-PG Resin and its Polychelates


positive bacteria) and S.aureus (gram positive bacteria), HEAP-1,2-PG resin


active.

For the inhibition of E.coli (gram negative bacteria), [La(III)-(HEAP-1,2-PG)]

(18 mm), [Pr(III)-(HEAP-1,2-PG)] (19 mm), [Nd(III)-(HEAP-1,2-PG)] (16 mm),

[Sm(III)-(HEAP-1,2-PG)] (17 mm), [Tb(III)-(HEAP-1,2-PG)] (16 mm ) and

[Dy(III)-(HEAP-1,2-PG)] (18 mm) were significantly active, while [Gd(III)-

(HEAP-1,2-PG)] (14 mm) was moderately active.

For the inhibition of B. substilis (gram positive bacteria), [La(III)-(HEAP-1,2-

PG)] (22 mm), [Pr(III)-(HEAP-1,2-PG)] (23 mm), [Nd(III)-(HEAP-1,2-PG)]

(20 mm), [Gd(III)-(HEAP-1,2-PG)] (19 mm), [Tb(III)-(HEAP-1,2-PG)] (20 mm)

and [Dy(III)-(HEAP-1,2-PG)] (22 mm) were significantly active, while

[Sm(III)-(HEAP-1,2-PG)] (15 mm) was moderately active.

For the inhibition of S.aureus (gram positive bacteria), [La(III)-(HEAP-1,2-

PG)] (22 mm), [Pr(III)-(HEAP-1,2-PG)] (18 mm), [Nd(III)-(HEAP-1,2-PG)]

(23 mm), [Sm(III)-(HEAP-1,2-PG)] (21 mm), [Gd(III)-(HEAP-1,2-PG)] (20 mm),

[Tb(III)-(HEAP-1,2-PG)] (19 mm) and [Dy(III)-(HEAP-1,2-PG)] (21 mm) were

significantly active.

For the inhibition of S. cerevisiae (yeast), [La(III)-(HEAP-1,2-PG)] (21 mm),

[Pr(III)-(HEAP-1,2-PG)] (22 mm), [Nd(III)-(HEAP-1,2-PG)] (19 mm), [Sm(III)-

(HEAP-1,2-PG)] (23 mm), [Gd(III)-(HEAP-1,2-PG)] (20 mm), [Tb(III)-(HEAP-1,2-

PG)] (23 mm) and [Dy(III)-(HEAP-1,2-PG)] (21 mm) were significantly active.



Table 5.2. Antimicrobial Activity data of the Polymeric Ligand

(HEAP-1,2-PG) and its Polychelates



coli

B.

substilis

S.

aureus

S.

cerevisiae

(HEAP-1,2-PG)n

-- -- -- 12

{[La(HEAP-1,2-PG)2(H

2O)

2}.OH]

n 18 22 22 21

{[Pr(HEAP-1,2-PG)2(H

2O)

2}.OH]

n 19 23 18 22

{[Nd(HEAP-1,2-PG)2(H

2O)

2}.OH]

n 16 20 23 19

{[Sm(HEAP-1,2-PG)2(H

2O)

2}.OH]

n 17 15 21 23

{[Gd(HEAP-1,2-PG)2(H

2O)

2}.OH]

n 14 19 20 20

{[Tb(HEAP-1,2-PG)2(H

2O)

2}.OH]

n 16 20 19 23

{[Dy(HEAP-1,2-PG)2(H

2O)

2}.OH]

n 18 22 21 21

DMSOb -- -- -- --





5.7.3. Antimicrobial Activity of HEAP-1,3-PD Resin and its Polychelates


positive bacteria) and S.aureus (gram positive bacteria), HEAP-1,3-PD resin


active.

For the inhibition of E.coli (gram negative bacteria), [La(III)-(HEAP-1,3-PD)]

(19 mm), [Pr(III)-(HEAP-1,3-PD)] (18 mm), [Nd(III)-(HEAP-1,3-PD)] (17 mm),

[Sm(III)-(HEAP-1,3-PD)] (18 mm), [Gd(III)-(HEAP-1,3-PD)] (17 mm), [Tb(III)-

(HEAP-1,3-PD)] (16 mm) and [Dy(III)-(HEAP-1,3-PD)] (18 mm) were



PD)] (20 mm), [Pr(III)-(HEAP-1,3-PD)] (19 mm), [Nd(III)-(HEAP-1,3-PD)]

(18 mm), [Sm(III)-(HEAP-1,3-PD)] (17 mm), [Gd(III)-(HEAP-1,3-PD)] (18 mm)

and [Tb(III)-(HEAP-1,3-PD)] (19 mm) were significantly active, while

[Dy(III)-(HEAP-1,3-PD)] (14 mm) was moderately active.


PD)] (21 mm), [Pr(III)-(HEAP-1,3-PD)] (20 mm), [Nd(III)-(HEAP-1,3-PD)]

(20 mm), [Sm(III)-(HEAP-1,3-PD)] (21 mm), [Gd(III)-(HEAP-1,3-PD)] (23 mm),

[Tb(III)-(HEAP-1,3-PD)] (19 mm) and [Dy(III)-(HEAP-1,3-PD)] (21 mm) were


For the inhibition of S. cerevisiae (yeast), [La(III)-(HEAP-1,3-PD)] (22 mm),

[Pr(III)-(HEAP-1,3-PD)] (21 mm), [Nd(III)-(HEAP-1,3-PD)] (23 mm), [Sm(III)-

(HEAP-1,3-PD)] (22 mm), [Gd(III)-(HEAP-1,3-PD)] (21 mm), [Tb(III)-(HEAP-1,3-

PD)] (22 mm) and [Dy(III)-(HEAP-1,3-PD)] (23 mm) were significantly active.




(HEAP-1,3-PD) and its Polychelates



coli

B.

substilis

S.

aureus

S.

cerevisiae

(HEAP-1,3-PD)n

-- -- -- 12

{[La(HEAP-1,3-PD)2(H

2O)

2}.OH]

n 19 20 21 22

{[Pr(HEAP-1,3-PD)2(H

2O)

2}.OH]

n 18 19 20 21

{[Nd(HEAP-1,3-PD)2(H

2O)

2}.OH]

n 17 18 20 23

{[Sm(HEAP-1,3-PD)2(H

2O)

2}.OH]

n 18 17 21 22

{[Gd(HEAP-1,3-PD)2(H

2O)

2}.OH]

n 17 18 23 21

{[Tb(HEAP-1,3-PD)2(H

2O)

2}.OH]

n 16 19 19 22

{[Dy(HEAP-1,3-PD)2(H

2O)

2}.OH]

n 18 14 21 23

DMSOb -- -- -- --





5.7.4. Antimicrobial Activity of HEAP-1,3-BD Resin and its Polychelates


positive bacteria) and S.aureus (gram positive bacteria), HEAP-1,3-BD resin


active.

For the inhibition of E.coli (gram negative bacteria), [La(III)-(HEAP-1,3-BD)]

(17 mm), [Pr(III)-(HEAP-1,3-BD)] (18 mm), [Nd(III)-(HEAP-1,3-BD)] (17 mm),

[Sm(III)-(HEAP-1,3-BD)] (19 mm), [Gd(III)-(HEAP-1,3-BD)] (16 mm) and

[Dy(III)-(HEAP-1,3-BD)] (16 mm) were significantly active, while [Tb(III)-

(HEAP-1,3-BD)] (15 mm) was moderately active.


BD)] (21 mm), [Pr(III)-(HEAP-1,3-BD)] (22 mm), [Nd(III)-(HEAP-1,3-BD)]

(19 mm), [Sm(III)-(HEAP-1,3-BD)] (20 mm), [Gd(III)-(HEAP-1,3-BD)] (18 mm),

[Tb(III)-(HEAP-1,3-BD)] (19 mm) and [Dy(III)-(HEAP-1,3-BD)] (23 mm) were



BD)] (20 mm), [Nd(III)-(HEAP-1,3-BD)] (21 mm), [Sm(III)-(HEAP-1,3-BD)]

(22 mm), [Gd(III)-(HEAP-1,3-BD)] (19 mm), [Tb(III)-(HEAP-1,3-BD)] (21 mm)

and [Dy(III)-(HEAP-1,3-BD)] (22 mm) were significantly active, while [Pr(III)-

(HEAP-1,3-BD)] (14 mm) was moderately active..

For the inhibition of S. cerevisiae (yeast), [La(III)-(HEAP-1,3-BD)] (23 mm),

[Pr(III)-(HEAP-1,3-BD)] (21 mm), [Nd(III)-(HEAP-1,3-BD)] (22 mm), [Sm(III)-

(HEAP-1,3-BD)] (23 mm), [Gd(III)-(HEAP-1,3-BD)] (21 mm), [Tb(III)-(HEAP-

1,3-BD)] (22 mm) and [Dy(III)-(HEAP-1,3-BD)] (23 mm) were significantly

active.




(HEAP-1,3-BD) and Its Polymer-metal complexes



coli

B.

substilis

S.

aureus

S.

cerevisiae

(HEAP-1,3-BD)n

-- -- -- 10

{La(HEAP-1,3-BD)2(H

2O)

2].OH}

n 17 21 20 23

{Pr(HEAP-1,3-BD)2(H

2O)

2].OH}

n 18 22 14 21

{Nd(HEAP-1,3-BD)2(H

2O)

2].OH}

n 17 19 21 22

{Sm(HEAP-1,3-BD)2(H

2O)

2].OH}

n 19 20 22 23

{Gd(HEAP-1,3-BD)2(H

2O)

2].OH}

n 16 18 19 21

{Tb(HEAP-1,3-BD)2(H

2O)

2].OH}

n 15 21 21 22

{Dy(HEAP-1,3-BD)2(H

2O)

2].OH}

n 16 23 22 23

DMSOb -- -- -- --





5.7.5. Antimicrobial Activity of HEAP-1,4-BD Resin and its Polychelates


positive bacteria) and S.aureus (gram positive bacteria), HEAP-1,4-BD resin


active.

For the inhibition of E.coli (gram negative bacteria), [La(III)-(HEAP-1,4-BD)]

(17 mm), [Pr(III)-(HEAP-1,4-BD)] (16 mm), [Nd(III)-(HEAP-1,4-BD)] (18 mm),

[Sm(III)-(HEAP-1,4-BD)] (16 mm) and [Gd(III)-(HEAP-1,4-BD)] (17 mm), was

significantly active, while [Tb(III)-(HEAP-1,4-BD)] (15 mm) and [Dy(III)-

(HEAP-1,4-BD)] (13 mm) (15 mm) were moderately active.











For the inhibition of S. cerevisiae (yeast), [La(III)-(HEAP-1,4-BD)] (20 mm),

[Pr(III)-(HEAP-1,4-BD)] (19 mm), [Nd(III)-(HEAP-1,4-BD)] (21 mm), [Sm(III)-

(HEAP-1,4-BD)] (18 mm), [Gd(III)-(HEAP-1,4-BD)] (20 mm), [Tb(III)-(HEAP-

1,4-BD)] (23 mm) and [Dy(III)-(HEAP-1,4-BD)] (19 mm) were significantly

active.



Table 5.5. Antimicrobial Activity data of the Polymeric Ligand and its

Polychelates



coli

B.

substilis

S.

aureus

S.

cerevisiae

(HEAP-1,4-BD)n

-- -- -- 12

{La(HEAP-1,4-BD)2(H

2O)

2].OH}

n 17 21 20 20

{Pr(HEAP-1,4-BD)2(H

2O)

2].OH}

n 16 23 19 19

{Nd(HEAP-1,4-BD)2(H

2O)

2].OH}

n 18 17 21 21

{Sm(HEAP-1,4-BD)2(H

2O)

2].OH}

n 16 19 21 18

{Gd(HEAP-1,4-BD)2(H

2O)

2].OH}

n 17 18 22 20

{Tb(HEAP-1,4-BD)2(H

2O)

2].OH}

n 15 21 20 23

{Dy(HEAP-1,4-BD)2(H

2O)

2].OH}

n 18 22 21 19

DMSOb -- -- -- --





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Documents

5.1. INTRODUCTION Contamination by …shodhganga.inflibnet.ac.in/bitstream/10603/39782/11/11...ropiness a sticky, stringy consistency caused by bacterial production of long-chain polysaccharides