Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 161 (2016) 3943

Contents lists available at ScienceDirect

Spectrochimica Acta Part A: Molecular and Biomolecular

Spectroscopy
journal homepage: www.elsevier.com/locate/saa

Synthesis, spectral studies and biological evaluation of 2-aminonicotinic

acid metal complexes
Muhammad Nawaz a,, Muhammad Waseem Abbasi b, Soleiman Hisaindee c, Muhammad Javed Zaki b,
Hira Fatima Abbas d, Hu Mengting e, M.Arif Ahmed f
a

Department of Nano-Medicine, Institute for Research and Medical Consultations, University of Dammam, Dammam, Saudi Arabia
Department of Botany, University of Karachi, Karachi 75250, Pakistan
Department of Chemistry, United Arab Emirates University, Al-Ain 15551, United Arab Emirates
d
Department of Pharmacology, College of Medicine, University of Malaya, Kuala Lumpur, Malaysia
e
State Key Laboratory of Advanced Technology for Materials Synthesis and Processing, Wuhan University of Technology, Wuhan 430070, PR China
f
H.E.J. Research Institute of Chemistry, International Center for Chemical and Biological Sciences, University of Karachi, Karachi 75270, Pakistan
b
c

a r t i c l e

i n f o

Article history:
Received 30 September 2015
Accepted 25 February 2016
Available online 26 February 2016
Keywords:
2-Aminonicotinic acid
Metal complexes
Spectral studies
Biological activity

a b s t r a c t
We synthesized 2-aminonicotinic acid (2-ANA) complexes with metals such as Co(II), Fe(III), Ni(II), Mn(II),
Zn(II), Ag(I),Cr(III), Cd(II) and Cu(II) in aqueous media. The complexes were characterized and elucidated
using FT-IR, UVVis, a uorescence spectrophotometer and thermo gravimetric analysis (TGA). TGA data showed
that the stoichiometry of complexes was 1:2 metal/ligand except for Ag(I) and Mn(II) where the ratio was 1:1.
The metal complexes showed varied antibacterial, fungicidal and nematicidal activities. The silver and zinc complexes showed highest activity against Bacillus subtilis and Bacillus licheniformis respectively. Fusarium oxysporum
was highly susceptible to nickel and copper complexes whereas Macrophomina phaseolina was completely inert
to the complexes. The silver and cadmium complexes were effective against the root-knot nematode Meloidogyne
javanica.
2016 Elsevier B.V. All rights reserved.

1. Introduction
Metal-based compounds have received great attention in medicinal
chemistry and have been widely studied in modern medicine for the
diagnosis and treatment of different human malignancies. For instance,
metallo-pharmaceutical agents have been described for the treatment
of cancer, arthritis, manic depression, and for use as anti-microbial
and anti-parasitic agents [1,2]. Pyridines have been extensively studied
for their biological and physiological activities. For example, nicotinic
acid has been assessed for its antibacterial properties [3] and as an
anti-hyperlipidemic agent raising HDL cholesterol level to reduce cardiovascular risks [4,5]. Nicotinamide and isonicotinamide were also
found to have an antifungal and antimicrobial activity [6]. Transition
metal complexes are widely used as potential therapeutics [7], they
are useful in health and skin care [8]. Many researchers reported biological activities of Cd complexes such as anti-cancer [9], cytotoxicity, antibacterial and antifungal activities [1012].
Nicotinic acid and its derivatives constitute important ligands in
coordination chemistry and have been extensively used as medicinal
agents. Morsy et al., [13] reported the preparation of silver complexes
Corresponding author.
E-mail address: nawwaz@gmail.com (M. Nawaz).

http://dx.doi.org/10.1016/j.saa.2016.02.022
1386-1425/ 2016 Elsevier B.V. All rights reserved.

with nicotinate ligands and studied their antibacterial activities against

antibiotic-resistant pathogens.
Soil-borne plant pathogens are equally important as they affect plant
cultures with economic, social and environmental implications. Many
soil-borne plant pathogens cause severe damage to different vegetable
crops and pulses by reducing crop yield and quality of bioproducts.
The soil-borne plant pathogens including nematodes, fungi and some
bacteria infect plant roots causing disruption of absorption and
translocation of nutrients and water from the soil resulting in mortality. Nematodes, including plant parasitic nematodes, are among
the most abundant macro and micro fauna of soil. Of these plant
parasitic nematodes, the root-knot nematodes (Meloidogyne spp.)
are the most destructive. They are widely distributed and reproduce
on over 2000 species of plants [14].
Root-knot nematodes (M. species) are obligate parasites and cause
root cells to become giant and multinucleated. Macrophomina phaseolina
is another pathogen that causes rotting of roots, stem and pods of hundreds of cultivated plant species [15].
M. phaseolina has a wide distribution and it can survive in soil and
dead plant debris for several years by forming sclerotia. Symptoms
including appearance of lesions on roots, stem and other parts of plants
which result in stunted growth, yellowing of leaves and reduction in
yield [16]. Similarly, Fusarium species are also very common plant

40

M. Nawaz et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 161 (2016) 3943

pathogens that cause root and stem rot and wilt diseases on a variety of
crop plants. Symptoms include yellowing of lower leaves and wilting.
In this work, we report the synthesis of metal complexes of
2-aminonicotinic acid (2-ANA). The complexes were characterized
by different spectroscopic techniques and their inhibitory effect
was evaluated against bacteria, fungi and nematode.

9 mm of internal diameter) after pouring Potato Dextrose agar (PDA).

About 10 mL of PDA was poured in each Petri plate and rotated well before solidication. A 6 mm pure culture disc was placed at the center of
the plate and incubated at 30 C for 47 days (depending upon tested
fungi). Fungal redial growth was measured in mm after incubation
and data of % inhibition by metal complexes were calculated.

2. Experimental

2.7. Nematicidal activity

2.1. Materials and chemicals

Nematicidal activity of complexes was assessed against the second

stage juveniles of root-knot nematodes, Meloidogyne javanica. Egg
masses were directly picked from roots with the help of ne needle
under a stereomicroscope and transferred into cavity blocks containing
sterilized distilled water. After hatching, the number of juveniles was
maintained around 2535 J2/mL. The metal complexes were diluted to
1000 ppm in the nematode suspensions. The number of dead juveniles
was counted after 24, 48 and 72 h of exposure. Cavity blocks without
metal complexes and with juveniles only were considered as controls.
Each treatment was repeated thrice. Nematodes were considered dead
if they did not move when probed with a ne needle [19]. Data of
dead juveniles were calculated and showed in mortality %.

All chemicals were of analytical reagent grade and were used without further purication. 2-Aminonicotinic acid, dimethyl sulfoxide
(DMSO) and metal salts Co(II), Fe(III), Ni(II), Mn(II), Zn(II),Ag(I),Cr(III),
Cd(II), and Cu(II) in the form of hydrated chloride or nitrate salts were
purchased from Sigma-Aldrich and BDH. Melting points of the complexes were determined on a Gallenkamp apparatus and reported as
uncorrected.
2.2. General procedure for the preparation of 2-aminonicotinic acid
(2-ANA) metal complexes
2 mmol (0.28 g) of 2-ANA and 2 mmol (0.11 g) of potassium hydroxide were stirred in deionized water for 30 min. To this mixture, an aqueous solution of metal salt (0.5 mmol) was added and the mixture was
stirred for 30 min. The precipitate formed was ltered, washed several
times with water, dried and stored in a silica gel desiccator.
2.3. Characterization
Infrared (IR) spectra of ligand and their metal complexes were recorded on a Nicolet Impact 410 FT-IR spectrometer (4000400 cm1)
as a potassium bromide (KBr) pellet with 4 cm1 resolution. UVVisible
spectra of compounds were recorded on a CARY 50 spectrophotometer
using dimethyl sulfoxide (DMSO) as a blank. Absorption and emission
spectra were recorded on a CARY 50 spectrouorometer in DMSO,
using a 10 mm quartz cell. Thermal analysis was carried out on a
Shimadzu thermo gravimetric analyzer (TGA). The sample (5.00 mg)
was heated in corundum crucibles up to 600 C, with a heating rate of
20 C min1 in air. The products of decomposition were calculated
from TG curves. The temperature ranges were determined by means
of a thermoanalyzer data processing module.
2.4. Biological cultures
Bacteria used for present study were isolated from the rhizosphere
of cultivated plants and identied as Bacillus subtilis (BSER) and Bacillus
licheniformis (BLF1). Fungi namely Fusarium oxysporum and M.phaseolina
were isolated from the soil of Department of Botany experimental eld,
University of Karachi. Root-knot nematodes were extracted from the
roots of egg plants maintained in culture.
2.5. Antibacterial activity
Antibacterial activity of metal complexes was tested by a disc diffusion technique [17]. Bacterial cells were spread on the surface of
LB (Lysogeny Broth) agar plate; prepared 6 mm diameter discs
were dipped in the solution of the complexes and placed on inoculated
agar surface. Plates were incubated for 24 h at 37 C in the dark, the
zone of growth inhibition was measured in millimeter around each of
the antibiotic disc, after incubation.
2.6. Fungicidal activity
Antifungal activity was carried out by an agar dilution method [18].
100 L of each metal complex was added to sterilize Petri plates (having

3. Results and discussion

3.1. Characterization
The physical properties and spectral data of the ligand (2-ANA) and
its metal complexes are presented in Table 1 and the proposed structure
is shown in Fig. 1. Most of the complexes were colored, stable in air and
soluble in DMSO. The proposed formulae of the complexes are in accordance with the elemental analysis presented in Table 2.
In order to nd out the coordination sites in the 2-ANA that may involve in complexation, the IR spectral data of 2-ANA and its metal complexes are compared and summarized in Table 1. The position and
intensities of the peaks are expected to change upon complexation.
The IR spectrum of the free ligand showed characteristic bands at
1709 and 3246 cm1 due to carboxylic and amino group stretching
modes respectively. In the complex spectra, the bands due to carboxylic
group were shifted to lower frequency as compared with the free ligand
suggesting that carboxylic group is coordinated to the metal ions. The
stretching band of the primary amine group also shifted to higher frequencies in most cases suggesting that the NH2 group is also involved
in coordination. The peaks are sharp indicating that there is no intra/
inter molecular hydrogen bonding in the solid state of the complexes.
Thus it indicates that the carboxylate oxygen and the primary amino
group of 2-ANA are chelated to the metal ions (Fig. 1). It is further
noted that the IR spectra of the complexes are similar indicating that
the coordination nature of the complexes is the same.
The UVVis absorption spectra of the ligand and metal complexes were recorded in water and DMSO respectively at 1.18 mM

Table 1
Analytical and physical data of 2-aminonicotinic acid and its metal complexes.
Complexes Formula

Color

Solubility m.p
(C)

max (cm1)
(nm)
C = O NH

ANA
1
2
3
4

White
Purple
Buff
Green
White

H2O
DMSO
DMSO
DMSO
DMSO

298 C
N300 C
N300 C
N300 C
N300 C

318
338
335
337
326

1709
1626
1705
1670
1624

3246
3404
3258
3437
3402

White
White
Green
White
Green

DMSO
DMSO
DMSO
DMSO
DMSO

N300 C
N300 C
N300 C
N300 C
N300 C

336
331
335
329
338

1629
1625
1707
1624
1702

3392
3427
3258
3398
3392

5
6
7
8
9

C6H6N2O2
Co(ANA)2
Fe(ANA)23H2O
Ni(ANA)2 H2O
Mn(ANA)
3H2O
Zn(ANA)2
Ag(ANA)
Cr(ANA)2H2O
Cd(ANA)2
Cu(ANA)2 H2O

M. Nawaz et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 161 (2016) 3943

41

Fig. 1. Proposed generalized structure of metalligand complex.

concentration. 2-ANA was found to be insoluble in DMSO and as a

result its UV spectrum was taken in water. The absorption spectra
of ligand and its complexes are shown in Fig. 2 and the data is summarized in Table 1. The spectrum of 2-ANA showed two bands at
max at 239 and 318 nm. When the ligand is complexed to the
metal, a bathochromic shift is observed in all cases in DMSO (compared to 2-ANA in water) suggesting that there may be an interaction between the ligand and the metal ions.
The uorescence spectra of 2-ANA and its metal complexes were
studied in water and DMSO at room temperature respectively. The
ligand solution was excited at 318 nm and an emission peak was
observed at 500 nm with very low intensity (Fig. 2). The solutions of
the complex spectra were excited at their respective absorption maxima (Table 1) and emission spectra were found around 380395 nm
(Fig. 3). Emission intensity of the metal complexes was higher than
that of the free ligand with an enhanced uorescence intensity of the
complexes. The latter is attributed to an increased in the rigidity of the
system attained through the coordination of 2-ANA to the metal ions,
thereby reducing energy loss by thermal vibrational decay [20].
The prepared complexes were subjected to thermal analysis and
a representative TGA curve corresponding to the decomposition of
metal complex 2 is shown in Fig. 4. The curve shows a step-wise degradation of the complex in the presence of air as temperature is increased
from room temperature to 600 C. Similar curves were observed for the
other complexes with variation in the number of steps. Table 3 summarized the decomposition steps of the complexes together with proposed
loss of entities during thermal treatment. These curves show mass
losses in two major stages. Generally, the rst stage corresponds to an
endothermic volatilization of physically adsorbed or coordinated
water molecules at approximately 200 C. The second stage, which
may comprise of two or more steps, involves the loss of organic residues
(in the form of CO2, H2O, and NOx) to give a metal oxide as a residue. The
TGA data were used to elucidate the structure of the different complexes
and we found that most of the complexes comprise of two ligands
(bidentate mode) bound to the metal ion except complexes 4 and 6
whose stoichiometry was 1:1 (monodentate mode). Also the complexes
had varied water of crystallization as shown in Table 3.

Fig. 2. Absorption spectra of ligand and its metal complexes.

3.2. Biological studies

Compound series 1 to 9 were tested for their anti-bactericidal activity against two Bacillus species, BSER and BLF1 at 500 and 1000 ppm
concentrations respectively. Greater zone of inhibition against BSER was
recorded when the media was inoculated with silver complex 6
(18.3 mm) at 1000 ppm followed by zinc complex 5 (16.6 mm)
when applied (P b 0.001) at the same concentration. The least activity against BSER was recorded against complex 9 (8.3 mm) at
1000 ppm. At 500 ppm, the inhibitory effects of the complexes are
generally less (351%) than that at the higher concentration. Nickel
complex 3 showed a considerable reduced activity (51%) when its
concentration is halved (Table 4) whereas the activity of copper
complex 9 is independent of concentration (0%).
Furthermore, zinc complex 5 gave greatest reduction of BLF1
by showing maximum zone of inhibition of 18.6 mm followed by
chromium complex 7 (17.0 mm) at 1000 ppm as compared to the control (Table 4). The least active was silver complex 6 (8.0 mm), which

Table 2
Elemental analysis data of metal complexes of 2-aminonicotinic acid.
Complexes

Formula

%Found (calcd.)
C

1
2
3
4
5
6
7
8
9

Co(ANA)2
Fe(ANA)23H2O
Ni(ANA)2H2O
Mn(ANA)3H2O
Zn(ANA)2
Ag(ANA)
Cr(ANA)2H2O
Cd(ANA)2
Cu(ANA)2 H2O

43.39 (43.26)
42.93 (43.03)
43.68 (43.56)
37.39(37.72)
42.99(42.69)
29.33 (29.54)
44.25 (44.45)
37.72(37.47)
42.78(42.93)

2.71(2.44)
2.69 (2.46)
2.83 (2.44)
2.56(2.11)
2.13(2.39)
1.43 (1.65)
2.29(2.49)
2.34 (2.10)
2.01(2.40)

16.48 (16.92)
17.18 (17.06)
16.76 (16.93)
14.43 (14.66)
16.26 (16.60)
11.26 (11.48)
17.54 (17.28)
14.72 (14.57)
16.28(16.69)

Fig. 3. Fluorescence spectra of ligand and its metal complexes.

42

M. Nawaz et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 161 (2016) 3943
Table 4
Antibacterial activity of metal complexes of 2-aminonicotinic acid.
Complexes

Zone of inhibition (mm)

BSER

BLF1

Concentration of complexes
1
2
3
4
5
6
7
8
9
Control (positive)
Control (negative)
LSD0.05 treatment
concentrations

Fig. 4. TGA curve of complex 2 (Fe(ANA)2.3H2O).

showed the best activity against BSER. At lower concentration (500 ppm),
the activities of the complexes were reduced and followed the same trend
as with the higher dose (Table 4).
Complexes used in this study demonstrated various degrees of
fungicidal activity towards F. oxysporum and M. phaseolina (Table 5).
Non-signicant reduction of F. oxysporum was achieved with all the
nine compounds tested at 500 and 1000 ppm concentrations respectively. Greater reduction of F. oxysporum colony was recorded when
media was supplemented with 1000 ppm of nickel complex 3 (overall
inhibition of 53%) followed by copper complex 9 (overall inhibition of
51%). Supplementation of media with only DMSO revealed very little effect on F. oxysporum colony. In general the activity of the complexes was

Table 3
Thermogravimetirc data of metal complexes of 2-aminonicotinic acid.
Complexes

Formula

Co(ANA)2

Fe(ANA)23H2O

Ni(ANA)2 H2O

Mn(ANA) 3H2O

Zn(ANA)2

Ag(ANA)

Cr(ANA)2H2O

Cd(ANA)2

Cu(ANA)2H2O

Steps

Temp.
range
(C)

Decomposed
assignment

Weight loss
found
(calcd.)%

1st
Residue
1st
2nd
3rd
Residue
1st
2nd
3rd
4th
Residue
1st
2nd
3rd
Residue
1st
2nd
Residue
1st
Residue
1st
2nd
3rd
Residue
1st
2nd
Residue
1st
2nd
3rd
Residue

380410

2 ANA

202280
315392
400534

3H2O
2 ANA

75.6 (77.5)
CoO
14.9 (14.0)
55.5 (51.0)

170254
254401
401437
453596

H2O
2 ANA

Fe2O3
6.5 (5.1)
73.5(77.5)

116158
181253
338435

3H2O
1 ANA

NiO
22.0 (21.8)
60.0 (63.2)

309451
456518

2 ANA

MnO
75.0 (76.1)

217301

1 ANA

ZnO
54.0 (53.6)

65101
181294
320389

H2O
2 ANA

7.5 (5.2)
79.0 (76.7)

304405
413541

2 ANA

Cr2O3
65 (67.0)

198268
270312
341480

H2O
2 ANA

CdO
7.0 (5.1)
70.5 (76.5)
CuO

500 ppm
1000 ppm
7.3
9.0
8.6
9.0
6.6
13.6
7.0
10.0
14.3
16.6
14.6
18.3
7.3
9.0
13.3
14.6
8.3
8.3
4.3 (disc dipped in DMSO)
0.0 (disc only)
2.3
1.0

500 ppm
1000 ppm
11.0
11.6
8.0
9.3
8.0
10.6
10.3
10.3
13.6
18.6
6.6
8.0
12.0
17.0
10.6
11.3
8.0
9.3
4.3 (disc dipped in DMSO)
0.0 (disc only)
2.8
1.2

diminished when the dose was reduced. However, least reduction was
accompanied by zinc complex 5 and iron complex 2 at 500 ppm concentration. M. phaseolina, on the other hand, do not show any responsive effect to the compounds with an average inhibition of 0%. The
fungistatic effect of compounds ranged between 19 to 53% (Table 5).
The complexes show selective inhibition of fungal growth and could
be useful in the control of one fungus over others when needed.
Nematicidal activity of isolated compounds was tested against
M. javanica second stage juveniles (J2) at different time intervals (24,
48, 72 h). All the complexes showed signicant (P b 0.001) nematicidal
activity. However, all juveniles were found to be dead when introduced
to silver complex 6 after 24 h interval, followed to cadmium complex 8
and zinc complex 5 also showed remarkable effect after 48 and 72 h exposure where nematicidal activity was 95 and 72% respectively (Table 6).
The lowest activity (13%) was recorded by copper complex 9 as compared
to water control which showed 0% mortality. Only DMSO solvent exhibited very least (3%) activity against M. javanica juveniles (Table 6). It
appears that the silver complex could be used as an effective control for
root-knot nematodes.
4. Conclusion
In summary, a series of metal complexes have been prepared
with 2-aminonicotinic acid ligand, involving 1:1 and 1:2 metal/ligand
stoichiometries. The synthesized metal complexes were characterized
by spectral and thermal studies. Thermal analysis of these metal complexes elucidated the composition, stoichiometry and also the number
Table 5
Fungicidal activity of metal complexes of 2-aminonicotinic acid.
Complexes

% Inhibition of fungi
M. phaseolina

F. oxysporum

Concentration of complexes
500 ppm
1
2
3
4
5
6
7
8
9
Control (positive)
Control (negative)
LSD0.05 treatment
concentrations

1000 ppm

0.0(disc dipped in DMSO)

0.0(disc only)

500 ppm

1000 ppm

24.4
37.8
20.7
27.0
40.7
53.0
29.6
34.8
19.3
31.1
43.7
47.8
24.0
30.0
41.8
46.3
44.5
51.5
7.4(disc dipped in DMSO)
8.1(disc only)
4.7
2.0

M. Nawaz et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 161 (2016) 3943
Table 6
Nematicidal activity of metal complexes of 2-aminonicotinic acid.
Complexes

1
2
3
4
5
6
7
8
9
Control (negative)
Control (positive)
LSD0.05

Time intervals (hours)

24

48

72

1.50
6.38
2.69
3.90
4.61
100.0
2.5
5.24
2.42
0
1.44
Treatment
time

5.11
34.78
27.08
5.24
28.43
100.0
12.81
82.17
12.55
0
2.08
3.64
1.90

22.44
46.57
68.90
26.60
71.59
100.0
26.40
95.18
12.70
0
3.34

of water crystallization. Furthermore these complexes exhibited

good biological activities against selected soil-borne bacteria, fungi
and nematodes.
References
[1] R. Bakhtiar, E.I. Ochiai, Gen. Pharmacol. 32 (1999) 525540.
[2] M. Navarro, Coord. Chem. Rev. 253 (2009) 16191626.
[3] W.L. McPheat, A.C. Wardlaw, P. Novotny, Infect. Immun. 41 (1983) 516522.

43

[4] E.T. Bodor, S. Offermanns, Br. J. Pharmacol. 153 (2007) S68S75.

[5] B.G. Brown, X.Q. Zhao, Am. J. Cardiol. 101 (2008) B58B62.
[6] T. Shimai, M.T. Islam, Y. Fukushi, Y. Hashidoko, R. Yokosawa, S.Z. Tahara,
Naturforsch. C 57 (2002) 323331.
[7] M.A. Ali, M.H. Kadir, M. Nazimuddin, S.M.M. Majumder, M.T.H. Tarafder, M.A. Khair,
Indian J. Chem. Sect A 27 (1988) 1064.
[8] C.D. Hufford, A.M. Clark, in: Atta-ur Rahman (Ed.), Studies in Natural Products
Chemistry, Elsevier, Amsterdam 1988, p. 421.
[9] Abdul Kareem, Hina Zafar, Asif Sherwani, Owais Mohammad, Tahir Ali Khan, J. Mol.
Struct. 1075 (2014) 1725.
[10] Foziah A. Al-Saif, Moamen S. Refat, Ten metal complexes of vitamin B3/niacin, J. Mol.
Struct. 1021 (2012) 4052.
[11] Hassan Keypour, Amir Hossein Jamshidi, Majid Rezaeivala, Laura Valencia, Polyhedron 52 (2013) 872878.
[12] T.A. Yousef, G.M. Abu El-Reash, M. Al-Jahdali, El-Bastawesy R. El-Rakhawy,
Spectrochim. Acta A Mol. Biomol. Spectrosc. 129 (2014) 163172.
[13] A.M. Morsy Abu-Youssef, R. Dey, Y. Gohar, A.A. Massoud, L. Ohrstrom, V. Langer,
Inorg. Chem. 46 (2007) 58935903.
[14] J.N. Sasser, D.W. Freckman, A world perspective on nematology: the role of the society, in: J.A. Veech, D.W. Dickson (Eds.), Vistas on Nematology, Society Of Nematologists, Inc., Hyattsville, MD 1987, pp. 714.
[15] O.D. Dhingra, J.B. Sinclair, Biology and pathology of Macrophomina phaseolina,
Universidade Federal de Viosa, Brazil, 1978 166.
[16] G.K. Gupta, S.K. Sharma, R. Ramteke, J. Phytopathol. 160 (2012) 167180.
[17] J.M. Swenson, J.B. Patel, J.K. Jorgensen, Special phenotypic methods for detecting
antibacterial resistance, Chapter 74, in: P.R. Murray, E.S. Baron, J.H. Jorgensen, M.L.
Landry, M.A. Pfaller (Eds.), Manual of Clinical Microbiology, nineth ed.ASM Press,
Washington DC 2007, pp. 11751176.
[18] L.A. Mitscher, R. Leu, M.S. Bathala, W. Wu, J.L. Beal, Lloydia 35 (1972) 157166.
[19] J.C. Cayrol, C. Djian, I. Pijarowski, Rev. Nematol. 12 (1989) 331336.
[20] S. Hisaindee, O. Zahid, M.A. Meetani, J.P. Graham, J. Fluoresc. 22 (2012) 677683.