MAKAUT

UNIVERSITY

Published on: ELSEVIER,2016 Submitted by:

Uttiya Dey, Soumendranath Ishika Das
Chatterjee, Naba kumar mondal M.Sc Microbiology
Guided by: Registration no :
Dr. Debdut Naskar 213001828110019
 TOPIC

Isolation and characterization of

arsenic-resistant bacteria and
possible application in
bioremediation.
 AIM

Isolation of bacteria which were resistant

to very high concentration of arsenic
forms , from contaminated water & are
able to reduce arsenic concentration.
 INTRODUCTION
•Arsenic is a toxic metalloid and it is present in the soil as
insoluble sulfides & sulfosalts.
•Arsenic contamination occurs mainly due to various
anthropogenic activities.
•Arsenic mainly exists in many forms in the environment,
among them Arsenite and Arsenate are abundant in nature.
•It can cause cellular damage, disrupting metabolic reaction,
and also effects on human health (when the concentration is
above 0.05ppm)
•Due to high magnitude of solubility , it’s not easy to remove
arsenic form contaminated water. Various conventional
method are used.
•Microbes can reduce toxicity by different mechanism.
MATERIALS & METHODS
•Study area
•Sampling
•Phsico-chemical analysis of water samples
•Isolation & enumeration of arsenic-resistant bacteria
•Phenotypic & biochemical characterization
•16SrDNA sequencing of the bacterial strains
•Bioremediation test
•Oxidation& reduction of arsenic by the isolates
RESULTS & DISCUSSION

Fig: 1 The study area is situated within 23^o 29’28.3”N to 23^0 31’54”N latitude and
88^0 17’ 4.3E to 88^0 21’ 56.1” longitude in purbasthali block of Burdwan
district.
Fig : 2 Scanning electron
micrograph of isolates.
(A) IsolateSW2 (control)
(B) isolate SW2 treated with
arsenite
(C) isolate SW4 (control)
(D) isolate SW4 treated with
arsenite
Both isolates are gram
positive and rod shaped.
•These isolates were positive for
catalase enzyme production but
unable for the production of
indole .
•Optimum growth occurred in
the media containing 4-6% of
sodium chloride
•Both isolates showed sensitivity
to recommended doses of some
antibiotics.
Fig : 3 Phylogenetic tree of (a) isolate SW2 & (b) isolate SW4
Significant removal of arsenate between SW2(53.29%) & SW4(50.37%)
was observed at 5% significance level from the media after 72h of
incubation.
Fig :5 Arsenite oxidation

(A: arsenite plate without bacteria, B: arsenite

plate inoculated with isolate SW2 & C: arsenite
plate inoculated with isolate SW4 ) .
After incubation, the presence of silver
arsenate in the media were observed. It was
observed that none of the bacteria has the
ability to reduce arsenate to arsenite , but both
of them can oxidize arsenite to arsenate.
 CONCLUSION
•It can be concluded that the two bacterial isolates ,
which were identifed as Bacillus sp. KM02 &
Aneurinibacillus aneurinilyticus strain BS1 can tolerate
arsenate concentration upto 4500 ppm & 550ppm of
arsenite concentration.
•They were halo tolerant.
•Both isolates have unique ability to oxidize arsenite to
less toxic arsenate.
•These two arsenic-resistant bacteria can be used as
novel pathway for the bioremediation of arsenic.
THANK
YOU!