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International Journal of Engineering & Technology, 7 (4.14) (2018) 121-126
International Journal of Engineering & Technology
Website: www.sciencepubco.com/index.php/IJET
Research paper
Biotransformation of Furaltadone, Furazolidone and
Nitrofurazone using Aspergillus Tamarii isolate TN-7 : in Vitro
Residual Identification and Quantification by HPLC-DAD
N S Mohammad
1
, M I A Halim
2
, M M Mahat
3
, M F Safian
2
, Z Z Ariffin
1*
1
School of Biology, Faculty of Applied Sciences, Universiti Teknologi MARA, 40450 Shah Alam, Selangor, Malaysia
2
School of Chemistry and Environment, Faculty of Applied Sciences, Universiti Teknologi MARA, 40450 Shah Alam, Selangor,
Malaysia
3
School of Physics and Material Studies, Faculty of Applied Sciences, Universiti Teknologi MARA, 40450 Shah Alam, Selangor,
Malaysia
*Corresponding author Email: drzaidah@salam.uitm.edu.my
Abstract
Nitrofurans (NFs) such as furaltadone (FTD), furazolidone (FZD) and nitrofurazone (NFZ) have been used as antibacterials and growth
promoters for the poultry and aquaculture industry. These antibiotics have now been banned from use due to their carcinogenic
properties; therefore there is an urgent need to remove or degrade NFs from contaminated areas. Aspergillus tamarii isolate TN-7
isolated from antibiotic overexposed soil shows an ability to degrade the NFs antibiotics. After 5 days of incubating of Aspergillus
tamarii isolate TN-7 with 500 μg/mL NF, the residual of the NF concentration was determined by High-Performance Liquid
Chromatography-diode array detector (HPLC-DAD). Solid phase extraction was performed to clean-up the fermentation broth prior to
HPLC-DAD analysis. Antimicrobial of the NFs residues showed a decreased in the percentage of inhibition that FTD, FZD and NFZ
were reduced to to 85.71 %, 75.86 % and 70.97 % after 96 hours of incubation. Quantification using HPLC-DAD showed, after 96 hours
of incubation, Aspergillus tamarii isolate TN-7 reduced furaltadone, furazolidone and nitrofurazone to 86.73 %, 37.49 % and 29.17 %
respectively. This finding shows that Aspergillus tamarii isolate TN-7 has the potential to be used as a bioremediation tool in removing
NF antibiotics from the contaminated areas.
Keywords: Aspergillus tamarii isolate TN-7; biotransformation; degradation; HPLC-DAD; nitrofuran.
1. Introduction
This Nitrofurans (NFs) such as furaltadone (FTD), furazolidone
(FZD) and nitrofurazone (NFZ) are widely used in veterinary
drugs for poultry and aquaculture. NFs are also used to treat
disease related to gastrointestinal tract and common disease by
bacteria and protozoa by inhibiting microbial enzymes that in-
volved in carbohydrate metabolism [1]. NFs have since been
banned and prohibited from usage in poultry or livestock
production. The U.S. Food Drug and Administration (FDA) ban
imports that have NF residues in them. The European Union (EU)
have banned the usage of NFs in livestock production since 1995
because of their carcinogenic and toxic effects on humans [2]. NF
parent drugs are unstable and are rapidly metabolized to their
metabolites (NFMs), FTD (AMOZ; 3-amino-5-morpholino-
methyl-2-oxazolidinone), FZD (AOZ; 3-amino-2-oxazolidinone)
and NFZ (SEM; semicarbazide). These NFMs maintain their
stability when bound to tissue or protein which can cause toxic
and carcinogenic effects. NFs administered on the poultry will be
excreted through faeces or urine of the chickens. These will enter
or will be introduced to soil
environment and cause soil contamination. According to Yu et al.,
[3], administration of NF in aquaculture will also cause the fish to
excrete the NFs or NFMs that then will enter the aquatic
environment and cause antibiotic contamination. Agriculture of
contaminated soil and aquatic environments exposed to these
antibiotics will then take up the NFs and NFMs, which will then
be transferred deleterious effects to humans. Continuous
deposition of antibiotics into the environment can increase the
resistance of bacteria towards these antibiotics. Thus, there is an
urgent need to degrade these antibiotics that have been deposited
into the environment. Aspergillus sp. is a species of fungus that
can be commonly found in soil [4] that showed great potential in
degrading pollutant. Aspergillus tamarii were reported to have the
ability to degrade endosulfan sulfate [5] and dyes [6]. The aim of
this study is to determine the ability of the Aspergillus tamarii
isolate TN-7 to degrade nitrofuran antibiotics.
2. Experimental
2.1 Reagents and chemicals
Analytical grade ethyl acetate (EtOAc > 99.95 %), was purchased
from Bendosen, HPLC grade methanol (MeOH > 99.90 %) from
Systerm, water HPLC grade was purchased from Amresco, and
acetonitrile (ACN > 99.90 %) was purchased from Merck.
Nitrofurans antibiotics, furaltadone (C
13
H
16
N
4
O
6
), furazolidone
(C
8
H
7
N
3
O
5
) and nitrofurazone (C
6
H
6
N
4
O
4
) were purchased from
Sigma-Aldrich.