Download Isolation of Halophilic Microorganisms From salted soil in Jazan area

Isolation of Halophilic Microorganisms From
salted soil in Jazan area
Thabet Ibrahim Qassem (200911926)
Biology Department -Faculty of Science- Jazan University
Abstract
Halophiles are a group of microorganisms that live in saline
environments and in many cases require salinity to survive.
Halophiles include a great diversity of organisms. At different
concentrations of Na Cl , 0,5,10 and 15% (w/v) bacteria and
fungi were isolated. The colony counts were increased at 5%
(w/v) and then decreased with increasing salt concentration up
to 15% (w/v). Different fungal species were isolate and
identified at different under different concentrations of NaCl.
Aspergillus sp, Penicillium sp, Cladosporium sp, Rhizopus sp
and Fusarium sp were detected undre different concentration
Key
words:
Halophilic
,
Microorganisms,
Soil
Introduction
Nearly 40% of world’s surface has salinity (Jadhav et al.,
2010). Salinization of soil is a serious problem and is
increasing gradually in many parts of the world, particularly in
arid and semiarid areas. At present, out of 1.5 billion hectares
of cultivated land around the world, about 77 million hectares
is affected by excess salt content (Evelin et al., 2009, Moradi et
al., 2011). At high salinity level, it was found that treatments
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supplied by biofertilization with yeast decreased the adverse
effect of salinity. Halophilic microorganisms are already in use
for some biotechnological processes, such as commercial
production of carotene, polymers, enzymes, compatible solutes
(Jadhav et al., 2010).
Kushner and Kamekura (1988) defined several categories of
micro-organisms on the basis of their optimal growth: (1)nonhalophiles are those that grow best in media containing less
than 0.2 M NaCl (1% salt). (2) slight halophiles grow best in
media with 0.2 to 0.5 M NaCl (1-3% salt). (3) moderate
halophiles grow best with 0.5 to 2.5 M NaCl (3-15% salt). (4)
extreme halophiles show optimal growth in media containing
2.5 to 5.2 M NaCl (15-32% salt) (Ventosa, 2006). Ibekewe et
al. (2010) reported that salinity and pH caused severe decrease
in the rhizosphere bacterial population. The unfavorable effects
of salinity on soil fertility are numerous. Its effects on uptake of
nutrients, absorption of moisture and soil structure are well
known. So we can use different halotolerant or halophile
bacteria for production of biofertilizer for optimum use of
saline land potential in agriculture. Halophiles are a group of
microorganisms that live in saline environments and in many
cases require salinity to survive. Halophiles include a great
diversity of organisms, like moderately halophilic aerobic
2
bacteria,
cyanobacteria,
sulphur-oxidizing
bacteria,
heterotrophic bacteria, anaerobic bacteria, archaea, protozoa,
fungi, algae and multicellular eukaryotes. Microorganisms that
are able to grow in the absence as well as in the presence of salt
are designated as halotolerant and those that are able to grow
above approximately 15% (w/v) NaCl (2.5 M) are considered
extremely halotolerant (Kushner et al., 1998, DasSarma and
Halophiles, 2001). According to Kushner (1978), many marine
organisms are slight halophiles (with 3% w/v NaCl in sea
water). Moderate halophiles optimally grow at 3-15% w/v Na
Cl, extreme halophiles at 25% w/v Na Cl.
Compatible solutes are low-molecular weight osmoregulatory
compounds which are highly water-soluble sugars, alcohols,
amino acids, betaines, ectoines or their derivatives. In addition
to their stabilizing effects, they are used as salt antagonists,
stress protective agents, moisturizers and therapeutics. They
stabilize enzymes, DNA and whole cells against stresses such
as freezing, drying and heating. They increase freshness of
foods by stabilizing components. Induction of osmolytes in
cells can increase protein folding and thereby improve salt
tolerance which could be useful in agriculture and xeriscaping
(Roberts
2005 and Detkova et al.,
2007). Halophilic
microorganisms usually adopt either of the two strategies of
3
survival in saline environments: ‘compatible solute’ strategy
and ‘salt-in’ strategy (Ventosa et al., 1998). The work aimed to
isolate halophilic or halotolerant microorganisms from salt soil.
Material and Methods
Microbial Analysis:
Salted soil samples from Jazan were
taken with the help of sterilized spatula at a depth of 5-15 cm .
One gm of soil was mixed with 9 ml of sterilized water and
mixed thoroughly. 1 ml from the solution was then mixed in 9
-2
ml sterilized water to make 10 dilution of this solution and in
-7
the same pattern dilutions up to 10 were prepared to determine
the microbial count.
The total numbers of microorganisms (
bacteria and fungi ) was counted. The numbers of colony
forming units (CFU) in the selective media were determined
by means of the serial dilution technique and the spread plate
method. Analyses were performed in three replicates. Viable
counts for fungi were performed using Dox agar medium.
Streptomycin
was
added
after
autoclaving
at
final
concentration of 30 mg ml. The plates were incubated at 28°C
and colonies were counted after 8 days . Viable counts for
bacteria was determined using a Nutrient agar medium
contained (per liter of water) Peptone ,5.0 g, Beef extract, 3.0g
Sodium chloride 5.0g, agar 20.0g.
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Results and Discussion
Halophilic and halotolerant microorganisms were studied with
many authors (Janda-Ulfig et al., 2009 , Soontharapirakkul and
Incharoensakdi 2010 , Shivanand and Mugeraya 2011). In our
study, numbers of bacteria and fungi ( CFU - colony forming
units ) increased with increasing NaCl concentration up to 5%
(w/v) and then decreased with increasin concentration
.Bacterial colonies not apperead at high concentration 15 % (
w/v) this results agree with obtained result by Ibekwe et al.
(2010). On the other hand , six fungal colonies apperead at high
concentration 15 % ( w/v) ( Table 1 & Fig1). Basically, the
effect of Na+ on the growth of different species of
microorganisms will differ due to growing water activity of
each
microorganism.
Microorganisms
under
hypertonic
environments either die or remain dormant except halotolerant
and
halophilic
microorganisms
(Chookietwattana,
2003).Different fungal species were isolate and identified at
different under different concentrations of NaCl. Aspergillus
sp, Penicillium sp, Cladosporium sp, Rhizopus sp and
Fusarium sp were detected undre different concentration (
Table 2).
5
Table (1) Numbers of Bacteria and Fungi ( CFU - colony forming
units ) X 103 / g dry salted soil at at different concentration of Na Cl
% ( w/v)
Na Cl Conc.
CFU - colony forming units of
% ( w/v)
Bacteria
Fungi
0
25
19
5
29
24
10
11
13
15
00
6
20
00
00
Fig (1) Numbers of fungi ( CFU - colony forming units ) X 103 / g dry
salted soil at at different concentration of Na Cl % ( w/v).
Fig (2) Numbers of bacteria ( CFU - colony forming units ) X 103 / g dry
salted soil at at different concentration of Na Cl % ( w/v).
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Table (2) Isolated fungus under different concentrations of NaCl
Fungus growth
Na Cl concentration ( w/v)
0
5
10
15
Aspergillus sp
+
+
+
Penicillium sp
+
+
+
+
Cladosporium sp
+
+
+
+
Rhizopus sp
+
+
Fusarium sp
+
+
Unknown species
+
+
+
+
Acknowledgements
We would like to thank Dr. Tarek M.Abd-El-Ghany for their
support ,valuable comments to improve the manuscript.
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‫عزل الكائنات الدقيقة المحبة للملوحة من تربة ملحية بمنطقة جازان‬
‫ثابت ابراهيم قاسم (‪)099099002‬‬
‫قسم األحياء – كلية العلوم – جامعة جازان‬
‫الملخص العربي‬
‫تم عمل تعداد كلي للكائنات الدقيقة البكتريا والفطريات من تربة ملحية في منطقة‬
‫جازان عند تركيزات مختلفة من ملح كلوريد الصوديوم (‪% 05،51،5105،0‬‬
‫وزن ‪/‬حجم) فوجد ان عدد المستعمرات الميكروبية تزداد عند تركيز ‪(%،‬وزن‬
‫‪/‬حجم) ولكن تقل بزيادة التركيز ‪ 5‬في نفس الوقت ظهرت مستعمرات فطرية عند‬
‫تركيز ‪ %1،‬بينما فشلت في النمو عند تركيز ‪ 5 % ،0‬علي الجانب اآلخر لم‬
‫تظهر مستعمرات بكتيرية عند تركيز ‪ .% 10‬وفي تلك الدراسة أيضا ً تم عزل‬
‫وتعريف الفطر يات النامية عند تركيزات مختلفة من الملح وذلك علي مستوي‬
‫الجنس‬
‫ووجد انها تنتمي لجنس االسبرجيللس والبنيسيليوم‬
‫والريزوباس‬
‫والفيزاريوم والكالدوسبوريم ‪5‬انواع اخري لم يتم التعرف عليها ‪ 5‬ووجد نوع من‬
‫فطر البنيسيليوم ينمو عند التركيزات المختلفة من الملح حتي ‪ ( % 1،‬وزن ‪/‬‬
‫حجم)‪.‬‬
‫‪11‬‬