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Romanian Biotechnological Letters Vol. 16, No. 4, 2011<br />

Copyright © 2011 University <strong>of</strong> Bucharest<br />

Pr<strong>in</strong>ted <strong>in</strong> Romania. All rights reserved<br />

ORIGINAL PAPER<br />

<strong>Investigation</strong> <strong>of</strong> <strong>Aspergillus</strong> <strong>niger</strong> <strong>growth</strong> <strong>and</strong> <strong>activity</strong> <strong>in</strong> a static magnetic<br />

flux density field<br />

Abstract<br />

6364<br />

Received for publication, August 12, 2010<br />

Accepted, July 14, 2011<br />

C. MATEESCU, N. BURUNŢEA, N. STANCU<br />

National Institute for Research <strong>and</strong> Development <strong>in</strong> Electrical Eng<strong>in</strong>eer<strong>in</strong>g ICPE-CA,<br />

313 Splaiul Unirii Str.., Bucharest-3, 030138, Romania, Tel: +40 213468297/ext. 115,<br />

Fax: +40 213468299, e-mail: carmen.mateescu@icpe-ca.ro<br />

Researches focused on the biodegradation <strong>of</strong> materials have <strong>in</strong>dicated that <strong>Aspergillus</strong> <strong>niger</strong><br />

microscopic fungus is destructor <strong>of</strong> electrical <strong>in</strong>sulations <strong>and</strong> other materials used <strong>in</strong> electrical <strong>in</strong>dustry<br />

<strong>in</strong> all the climatic zones. Static magnetic fields can exert some <strong>in</strong>fluence over microorganisms <strong>and</strong> this<br />

effect depends on many factors such as type <strong>and</strong> magnitude <strong>of</strong> magnetic field, type <strong>of</strong> microorganism,<br />

temperature, length/duration <strong>of</strong> exposure, <strong>growth</strong> media etc.<br />

The aim <strong>of</strong> the present study was to <strong>in</strong>vestigate the effect <strong>of</strong> the static magnetic <strong>in</strong>duction field<br />

on the <strong>growth</strong> <strong>and</strong> metabolic <strong>activity</strong> <strong>of</strong> <strong>Aspergillus</strong> <strong>niger</strong> fungus. The results <strong>of</strong> the study will give a<br />

better underst<strong>and</strong><strong>in</strong>g <strong>of</strong> biodegradation processes which occur on materials <strong>and</strong> electrical equipments.<br />

We <strong>in</strong>vestigated the <strong>growth</strong> <strong>of</strong> the fungus developed on nutritious Czapek-Dox solid media when<br />

exposed to a magnetic field <strong>of</strong> 0.50 T <strong>and</strong> 0.62 T respectively, for a period <strong>of</strong> 7 days. The exposed<br />

results were compared with those <strong>of</strong> sham controls, where the fungus developed <strong>in</strong> the same conditions<br />

as the exposed ones but without apply<strong>in</strong>g the magnetic field exposure. In order to assess the grow<strong>in</strong>g<br />

<strong>and</strong> morphology <strong>of</strong> the fungus colonies, the cultures have been macroscopically exam<strong>in</strong>ed after 48, 72,<br />

96 <strong>and</strong> 168 hours respectively, as well as microscopically at the end <strong>of</strong> the total exposure period.<br />

Keywords: fungus, biodegradation, static magnetic field, electrical equipment<br />

Introduction<br />

Whereas alternat<strong>in</strong>g current (AC) transmission l<strong>in</strong>es produce AC (mostly 50 Hz)<br />

electric <strong>and</strong> magnetic fields, static electric <strong>and</strong> magnetic fields are produced by direct current<br />

(DC) transmission l<strong>in</strong>es which is an economical way for power distribution over long<br />

distances. Modern magnetic levitation (maglev) systems use magnetic flux densities around 1<br />

T directly on the rails [1]. It has to be noticed that AC as well as DC electric fields are easily<br />

reduced by all k<strong>in</strong>d <strong>of</strong> obstacles <strong>and</strong> materials whereas the magnetic field not <strong>and</strong> is pass<strong>in</strong>g<br />

through build<strong>in</strong>gs, humans, <strong>and</strong> most <strong>of</strong> the materials.<br />

S<strong>in</strong>ce AC <strong>and</strong> DC magnetic fields are most likely to penetrate the body, they are the<br />

component <strong>of</strong> electromagnetic fields that are usually studied <strong>in</strong> relation to the effects on liv<strong>in</strong>g<br />

systems.<br />

It is well known that mould might cause damage to materials <strong>and</strong> equipments <strong>and</strong><br />

create health problems <strong>in</strong> warm <strong>and</strong> humid environments. Dur<strong>in</strong>g the normal <strong>growth</strong> <strong>of</strong> fungi,<br />

there is an enzymatic secretion that accelerates a chemical transformation <strong>of</strong> organic<br />

substances such as <strong>in</strong>sulation materials, wood derivates (paper), some types <strong>of</strong> seals, coat<strong>in</strong>gs,<br />

as well as many other types <strong>of</strong> materials from the structure <strong>of</strong> the electrical equipments. Many<br />

m<strong>in</strong>erals are also destroyed by the fungus.<br />

<strong>Aspergillus</strong> <strong>niger</strong> is a fungus so widely distributed that it is ubiquitous <strong>in</strong> nature. This<br />

fungus can spread easily <strong>and</strong> colonize a wide range <strong>of</strong> substrates, <strong>in</strong>clud<strong>in</strong>g wood <strong>and</strong> plastics<br />

<strong>in</strong> the components <strong>of</strong> many devices <strong>and</strong> equipments. <strong>Aspergillus</strong> <strong>niger</strong> fungus may also be<br />

found <strong>in</strong> compost <strong>and</strong> other decay<strong>in</strong>g organic material.


C. MATEESCU, N. BURUNŢEA, N. STANCU<br />

This paper aimed to <strong>in</strong>vestigate the <strong>growth</strong> <strong>and</strong> <strong>activity</strong> <strong>of</strong> <strong>Aspergillus</strong> <strong>niger</strong> fungus by<br />

expos<strong>in</strong>g it under two magnetic flux densities (B-field), for a total exposure period <strong>of</strong> 168<br />

hours. S<strong>in</strong>ce the present study is quite orig<strong>in</strong>al data on a similar issue are scarce <strong>in</strong> literature.<br />

Manoliu et al (2007) [3] <strong>in</strong>vestigated the development <strong>of</strong> <strong>Aspergillus</strong> <strong>niger</strong> exposed to<br />

a static B-field vary<strong>in</strong>g from 40-80 T. The results showed a 1.5 - 2 times faster <strong>growth</strong> rate<br />

than <strong>in</strong> sham controls. Moreover, it has been showed that the B-field exposure can have an<br />

effect on the biodegradability <strong>of</strong> materials by enhanc<strong>in</strong>g the <strong>growth</strong> rate <strong>and</strong> the<br />

aggressiveness <strong>of</strong> the fungus. It has also been noticed that a B-field exposure <strong>of</strong> more than<br />

350 T may delay the <strong>growth</strong> <strong>of</strong> microscopic fungus.<br />

The <strong>in</strong>fluence <strong>of</strong> the B-field exposure on cellulases <strong>and</strong> catalase <strong>activity</strong> <strong>in</strong> cellulolytic<br />

fungi Chaetomium globosum <strong>and</strong> Trichoderma viride cultivated on media with waste from<br />

<strong>in</strong>dustry <strong>of</strong> panification has been <strong>in</strong>vestigated too. It has been shown that the <strong>activity</strong> <strong>of</strong> these<br />

enzymes was <strong>in</strong>fluenced by both studied cellulolytic species <strong>and</strong> the exposure length to the B-<br />

field [4].<br />

Besides <strong>Aspergillus</strong> <strong>niger</strong> fungus, the effect <strong>of</strong> static magnetic fields has also been<br />

studied at the molecular <strong>and</strong> cellular levels <strong>in</strong> bacteria species, yeasts <strong>and</strong> molds. Moreover,<br />

the <strong>growth</strong> rate, colony size, gas production, viability <strong>and</strong> mutation effects, enzyme <strong>activity</strong><br />

<strong>and</strong> germ<strong>in</strong>ation <strong>of</strong> spores [4] have also been <strong>in</strong>vestigated.<br />

The motivation <strong>of</strong> our study is double. Firstly, because the <strong>Aspergillus</strong> <strong>niger</strong> fungus is<br />

widely spread on electrical equipment <strong>in</strong> various environmental conditions <strong>and</strong> secondly<br />

because there is a lack <strong>of</strong> scientific data on the effects <strong>of</strong> static magnetic fields on this fungus.<br />

Materials <strong>and</strong> Methods<br />

For <strong>Aspergillus</strong> <strong>niger</strong> <strong>growth</strong> it has been used Czapek-Dox agar, a synthetic solid<br />

medium, conta<strong>in</strong><strong>in</strong>g sucrose as carbon source <strong>and</strong> nitrate as nitrogen source. On Czapek-Dox<br />

agar, colonies <strong>of</strong> <strong>Aspergillus</strong> <strong>niger</strong> consist <strong>of</strong> a compact white or yellow basal felt covered by<br />

a dense layer <strong>of</strong> dark-brown to black colonial heads.<br />

The culture medium was prepared by dissolv<strong>in</strong>g <strong>of</strong> the follow<strong>in</strong>g components <strong>in</strong>to one<br />

liter <strong>of</strong> distilled water: sucrose 30 g, sodium nitrate 3 g, potassium chloride 0.5 g, magnesium<br />

sulfate heptahydrate 0.5 g, iron (II) sulfate heptahydrate 0.01 g, di-potassium hydrogen<br />

phosphate 1 g <strong>and</strong> agar 15 g. The suspension was brought to the boil <strong>in</strong> order to dissolve<br />

completely all the <strong>in</strong>gredients <strong>and</strong> then was sterilized by autoclav<strong>in</strong>g at 121 0 C for 15 m<strong>in</strong>utes<br />

[5]. The molten medium was poured <strong>in</strong>to three 7 cm diameter Petri dishes.<br />

The plates were <strong>in</strong>oculated with <strong>Aspergillus</strong> <strong>niger</strong> us<strong>in</strong>g a sterile wire. The fungus<br />

which we used for test<strong>in</strong>g was 30 days old <strong>and</strong> was obta<strong>in</strong>ed from our own collection.<br />

The environment conditions for fungus <strong>growth</strong> were room temperature <strong>of</strong> 25 ± 2 0 C,<br />

natural light <strong>and</strong> the humidity provided by the culture medium [6].<br />

For study<strong>in</strong>g the efficiency <strong>of</strong> the B-field, one sham control <strong>and</strong> two irradiated<br />

(exposed) <strong>in</strong>oculation plates were used. Whereas the sham <strong>in</strong>oculated plate was treated <strong>in</strong> the<br />

same way as the irradiated ones but without apply<strong>in</strong>g the B-field, the two other <strong>in</strong>oculated plates<br />

were exposed to a B-field <strong>of</strong> 0.5 T <strong>and</strong> 0.62 T respectively. Though the total exposure time was<br />

168 hours, the cultures were periodically analyzed after 48, 72, 96 <strong>and</strong> 168 hours respectively.<br />

After <strong>in</strong>cubations all the cultures were microscopically analyzed as regards the<br />

<strong>Aspergillus</strong> <strong>niger</strong> sporulation <strong>growth</strong>, us<strong>in</strong>g an optical <strong>in</strong>verted microscope type Nikon<br />

Eclipse Ti-E fitted with a confocal system Eclipse C1si. The microscopic exam<strong>in</strong>ation <strong>of</strong> the<br />

fungus was performed <strong>in</strong> bright field with a magnitude <strong>of</strong> 40 X.<br />

In order to make a comparative analysis <strong>of</strong> <strong>Aspergillus</strong> <strong>niger</strong> <strong>growth</strong> under different<br />

static magnetic flux densities, two magnetic systems have been calculated <strong>and</strong> designed for<br />

Romanian Biotechnological Letters, Vol. 16, No. 4, 2011 6365


<strong>Investigation</strong> <strong>of</strong> <strong>Aspergillus</strong> <strong>niger</strong> <strong>growth</strong> <strong>and</strong> <strong>activity</strong> <strong>in</strong> a static magnetic flux density field<br />

perform<strong>in</strong>g exposures to B-field <strong>of</strong> 0.5 T <strong>and</strong> 0.62 T. These magnetic systems consisted <strong>of</strong> the<br />

follow<strong>in</strong>g components:<br />

a. Magnetic circuit made <strong>of</strong> steel type OL 35;<br />

b. Permanent magnets, made <strong>of</strong> magnetic alloy based on NdFeB, hav<strong>in</strong>g the follow<strong>in</strong>g<br />

magnetic field characteristics: Br = 1.2 T, Hcb = 970 kA/m, (BH) max = 35 MGsOe,<br />

respectively;<br />

c. Polar pieces made <strong>of</strong> steel type OL 35.<br />

The exposure strength <strong>of</strong> the B-field <strong>in</strong> air has been performed by simulation <strong>of</strong><br />

magnetic fields us<strong>in</strong>g a magnetic calculation s<strong>of</strong>t type FEM (F<strong>in</strong>it Element Method).<br />

Results <strong>and</strong> discussions<br />

For the <strong>in</strong>vestigations <strong>of</strong> the effect <strong>of</strong> a static B-field on the mycelial <strong>growth</strong> <strong>and</strong><br />

conidia formation, the plates were exam<strong>in</strong>ed after an <strong>in</strong>cubation period <strong>of</strong> 48, 72, 96 <strong>and</strong> 168<br />

hours respectively. Figures 1-4 (a,b,c) show the <strong>Aspergillus</strong> <strong>niger</strong> <strong>growth</strong> without B-field<br />

exposure (sham control), under a B-field exposure <strong>of</strong> 0.5 T <strong>and</strong> <strong>of</strong> 0.62 T, for 48, 72, 96 <strong>and</strong><br />

168 hours respectively.<br />

Dur<strong>in</strong>g the first 48 hours we observed that the non-exposed fungus has grown faster<br />

than the two other samples which were exposed to the B-field. It means that after an<br />

<strong>in</strong>cubation <strong>of</strong> 48 hours, the <strong>growth</strong> <strong>of</strong> <strong>Aspergillus</strong> <strong>niger</strong> was characterized by the development<br />

<strong>of</strong> small but compact colonies with dense sporulation, spread from the <strong>in</strong>oculated po<strong>in</strong>t to the<br />

whole surface <strong>of</strong> the culture medium (Figure 1 a). The fungus exposed to the B-field produced<br />

less but larger colonies, with strong sporulation. The colonies are not spread on the whole<br />

surface <strong>of</strong> the culture medium (Figures 1b, 1c).<br />

a b c<br />

Fig 1. (a,b,c) <strong>Aspergillus</strong> <strong>niger</strong> <strong>growth</strong> after 48 hours <strong>of</strong> <strong>in</strong>cubation (a -without magnetic field;<br />

b – magnetic field <strong>of</strong> 0.5T; c – magnetic field <strong>of</strong> 0.62T)<br />

After a B-field exposure <strong>of</strong> 72 hours, the mould followed an atypical <strong>growth</strong>,<br />

characterized by about 20 larger <strong>and</strong> bombastic colonies hav<strong>in</strong>g white-yellowish normal<br />

aspect <strong>and</strong> very few dark-brown colonial heads. A stronger B-field exposure resulted <strong>in</strong> a<br />

larger but rarer colony formation <strong>in</strong> the culture medium. Thus, for the plate exposed to a B-<br />

field <strong>of</strong> 0.5 T (Figure 2b), we observed more but smaller colonies which tend to jo<strong>in</strong> each<br />

other, compar<strong>in</strong>g to the sample exposed <strong>in</strong> B-field <strong>of</strong> 0.62 T (Figure 2c). By compar<strong>in</strong>g the<br />

0.5 T (Figure 2b) <strong>and</strong> the 0.62 T exposure results, we observe <strong>in</strong> the first case more but<br />

smaller colonies which tend to stick together than <strong>in</strong> the second case.<br />

6366 Romanian Biotechnological Letters, Vol. 16, No. 4, 2011


C. MATEESCU, N. BURUNŢEA, N. STANCU<br />

a b c<br />

Fig 2. (a,b,c) <strong>Aspergillus</strong> <strong>niger</strong> <strong>growth</strong> after 72 hours <strong>of</strong> <strong>in</strong>cubation (a -without magnetic field; b – magnetic<br />

field <strong>of</strong> 0.5; c – magnetic field <strong>of</strong> 0.62T)<br />

After 96 hours <strong>of</strong> <strong>in</strong>cubation, some notches or cuts became visible on the swollen<br />

surface <strong>of</strong> the colonies. They are more explicit for the stronger B-field <strong>of</strong> 0.62 T (Figure 3c).<br />

These notches did not arise when the B-field was not applied.<br />

a b c<br />

Fig 3. (a,b,c) <strong>Aspergillus</strong> <strong>niger</strong> <strong>growth</strong> after 96 hours <strong>of</strong> <strong>in</strong>cubation (a -without magnetic field;b – magnetic field<br />

<strong>of</strong> 0.5T; c – magnetic field <strong>of</strong> 0.62T)<br />

After an <strong>in</strong>cubation period <strong>of</strong> 168 hours (at the end <strong>of</strong> test<strong>in</strong>g), we observed a<br />

relatively homogenous <strong>growth</strong> <strong>of</strong> the fungus <strong>in</strong> the plate not exposed to the B-field (Figure<br />

4a). The colonies became completely black on the whole surface <strong>of</strong> the culture medium.<br />

The sample exposed to the strongest B-field <strong>of</strong> 0.62 T (Figure 4c) showed <strong>in</strong>tensively<br />

black colored <strong>and</strong> large colonies. In this case the black colonial heads are denser as compared<br />

to the fungus developed <strong>in</strong> cultures not exposed to a B-field . This atypical <strong>growth</strong> does not<br />

cover the entire surface <strong>of</strong> the Petri dish. The sample exposed <strong>in</strong> the B-field <strong>of</strong> 0.5 T (Figure<br />

4b) shows an <strong>in</strong>termediary aspect between the two plates.<br />

a b c<br />

Fig 4. (a,b,c) <strong>Aspergillus</strong> <strong>niger</strong> <strong>growth</strong> after 168 hours <strong>of</strong> <strong>in</strong>cubation (a -without magnetic field; b – magnetic<br />

field <strong>of</strong> 0.5T; c – magnetic field <strong>of</strong> 0.62T)<br />

Romanian Biotechnological Letters, Vol. 16, No. 4, 2011 6367


<strong>Investigation</strong> <strong>of</strong> <strong>Aspergillus</strong> <strong>niger</strong> <strong>growth</strong> <strong>and</strong> <strong>activity</strong> <strong>in</strong> a static magnetic flux density field<br />

Figure 5 shows a microscopic photograph <strong>of</strong> radiate conidial head <strong>and</strong> brown-colored<br />

round-shaped unicellular conidia <strong>of</strong> <strong>Aspergillus</strong> <strong>niger</strong>.<br />

Fig. 5. Micrograph <strong>of</strong> <strong>Aspergillus</strong> <strong>niger</strong> conidia with a full-field on the left <strong>and</strong> a detail shot on the right<br />

Conidia <strong>in</strong> cha<strong>in</strong>s or detached <strong>and</strong> dispersed can be observed <strong>in</strong> this microscopy<br />

image. S<strong>in</strong>gle or paired conidia may resemble yeast cells. The size <strong>of</strong> spores <strong>of</strong> <strong>Aspergillus</strong><br />

<strong>niger</strong> ranges from 2 to 4 microns.<br />

Conclusions<br />

The effect <strong>of</strong> a static magnetic flux density on the <strong>growth</strong> <strong>and</strong> <strong>activity</strong> <strong>of</strong> <strong>Aspergillus</strong><br />

<strong>niger</strong> have been macroscopically <strong>and</strong> microscopically <strong>in</strong>vestigated by means <strong>of</strong> two exposure<br />

strengths. It is concluded that the static magnetic field produces an atypical <strong>growth</strong> <strong>of</strong> the<br />

fungus that is characterized by less <strong>and</strong> swollen, bombastic colonies which did not spread on<br />

the entire surface <strong>of</strong> the culture medium. From the comparison between the exposed <strong>and</strong> the<br />

sham, we conclude that the magnetic field is an efficient <strong>in</strong>hibitor <strong>of</strong> the surface <strong>growth</strong> <strong>of</strong> the<br />

fungus.<br />

The validation or practical application <strong>of</strong> the present study lies <strong>in</strong> the fact that the<br />

results can be used for develop<strong>in</strong>g magnetic field sources/methods that are able to reduce or<br />

elim<strong>in</strong>ate bio-damage <strong>of</strong> components <strong>of</strong> electrical equipments or <strong>in</strong>stallations <strong>and</strong> materials<br />

which are sensitive to <strong>Aspergillus</strong> <strong>niger</strong> <strong>and</strong> perhaps to other fungi.<br />

References<br />

1. ICNIRP 13/2003. Exposure to Static <strong>and</strong> Low Frequency Electromagnetic Fields. Biological Effects <strong>and</strong><br />

Health Consequences (0-100 kHz). Publication <strong>of</strong> the International Commission on Non-Ionizng Radiation<br />

Protection. Pr<strong>in</strong>ted by Märkl-Druck, München.<br />

2. S. A. SEMENOV,KLARA Z. GUMARGALIEVA,GENNADIĬ EFREMOVICH ZAIKOV, Biodegradation <strong>and</strong><br />

durability <strong>of</strong> materials under the effect <strong>of</strong> microorganisms, VSP BV, ISBN 90-6764-388-2, pp. 190-192, (2003).<br />

3. A. MANOLIU, L. OPRICK, D. CREANGA, The <strong>in</strong>fluence <strong>of</strong> the static magnetic field (SMF) on some<br />

biochemical parameters <strong>in</strong> cellulolytic fungi Chaetomium globosum <strong>and</strong> Trichoderma viride cultivated on<br />

media supplemented with panification <strong>in</strong>dustrial wastes, Rom. Journal Biol, Vol. 51-52, 2007, pp. 25-37<br />

4. P.E KOVACS., R.L VALENTINE., P.J. ALVAREZ, The effect <strong>of</strong> static magnetic fields on biological<br />

systems: Implications for enhanced biodegradation, Crit. Rev. Environ. Sci. Technol., 27, 1997, pp. 319-382.<br />

5. www.sigmaaldrich.com/etc/medialib/docs/Fluka/usage/70185_czapek_dox_agar.Par.0001.File.tmp/70185_c<br />

zapek_dox_agar.pdf<br />

6. M. MITITIUC, C. TANASE, Micologie, Editura Univ. Al. I. Cuza, Iasi, 2000, pp. 47-52<br />

6368 Romanian Biotechnological Letters, Vol. 16, No. 4, 2011

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