2019, Vol. 2, No. 3, 279 – 288
http://dx.doi.org/10.11594/jk6em.xx.xx.xx
Antimicrobial Production by an Actinomycetes Isolated from the Termite Nest
Ni Putu Ratna Ayu Krishanti *, Deni Zulfiana, Bramantyo Wikantyoso
Research Center for Biomaterials, Indonesian Institute of Sciences, Bogor 16911, Indonesia Article history:
Submission August 2018 Revised August 2018 Accepted September 2018
ABSTRACT
Actinomycetes are Gram positive bacteria that have been detected in diverse ecological niches. Their member species are known to be a main source of vari- ous bioactive compounds. The discovery of Actinomycetes from diverse and un- explored resources has also been linked to increased opportunities to obtain novel bioactive compounds. Insect nest material is being investigated as a new source of novel antimicrobial producing Actinomycetes, which could be har- nessed for therapeutic potential. A total of 10 Actinomycetes isolates were col- lected from the nest of Nasutitermes sp. in Pananjung Pangandaran Nature Re- serve. These isolates were evaluated for antimicrobial activity against the chal- lenge bacteria (Eschericia coli, Staphylococcus aureus, Bacillus subtilis, Serra- tia marcescens) and fungi (Fomitopsis palustris, Fusarium oxysporum, Tricho- derma viridae) by dual culture method. The result revealed that several isolates were active against fungi and bacteria. Isolate Pn-TN2 showed the highest level of antibacterial inhibition and the highest antifungal inhibition with Inhibition Rate value more than 80%. By morphological and 16S rRNA gene sequence analysis strongly suggested that the isolate Pn-TN2 belonged to Streptomyces prasinopilosus. We suggested that termite nest is a potential source of bioactive strains of cultivable Actinomycetes for future biotechnological needs.
Keywords: Actinomycetes, antimicrobial, Nasutitermes sp., Streptomyces prasinopilosus, termite nest
*Corresponding author:
E-mail: [email protected] Email:
Introduction
Indonesia excels in a period of 2045 prepared by the application from the map of the industrial revolution ere 4.0. answering education as a solu- tion to the impact of the industrial era 4.0. re- quired entrepreneur education. The concept of entrepreneurial education will create creative and innovative children who able to overcome the ob- stacles in the future. Moravec states that industry 4.0 requires the education to build individual innovation practices and student empowerment teams to produce innovation (Sarikaya &
Coskun, 2019). Schools are educational institutions designed for the development of human potentials. Also, the purpose of education is to improve the quality of human resources with the anticipation of producing superior and characterized human beings, raised through the
affirmation of each activity that is integrated into entrepreneurship-based learning in school.
Ningrum's opinion (2017) reported entrepreneurship as a soul, attitude, ability to create something new, valuable and useful for personal consumption and others, while entrepreneurs are skilled personnel in the aspect of developing personal businesses and taking advantage of opportunities, to improve their lives. Moreover, Nurseto (2010) established that efforts needed to instill the required entrepreneurship for teachers ought not to be limited to master the concepts, but also have the ability to perform the related activities. These include the application of all existing conceptions.
The plans towards learning conducted at Mata Najwa Kindergarten which involved the application of entrepreneurship activities in early
How to cite:
Research Article
NPRA Krishanti, D Zulfiana, B Wikantyoso et al., 2018 / Antimicrobial Production by an Actinomycetes Isolate
childhood, including several learning components, encompassing: 1) the objectives; 2) materials; 3) educators; 4) students; and 5) strategies (Sofino, 2017). This is followed by the inculcation of entrepreneurial values, performed in a series of activities that are initiated with planning, learning, implementing and evaluating.
Sani and Yunus (2018) in Malaysia showed the need to relate the results of research on teaching design or planning to one another and not to a specific subject. Furthermore, the information required is centered on the objective learning content, approaches, methods used, the imparting techniques and assessment design of the teachers’
teaching and learning outcomes. These are needed to play the role of an evaluation material, which ought to be performed and added on instances where there are deficiencies.
the cooking class is innovative programs i n preschool children. These are usually con- ductedto promote healthy eating behaviors,at- tained by providingthe knowledge on basic nutr ition, food selection, menu planning, time-sav ing tips in the kitchen and food preparation s kills to parents and caregivers (Condrasky, Graham & Kamp, 2006).
This is also significantly increased their willin gness to explore fruits and vegetables, as rep orted by parents. Also, the effectiveness of co oking on children’s food-related preferences, attitude and behaviors is promoted (Hersch et a l., 2014). The results and discussions stipulated that the values of entrepreneurship need to be internalized in students include independent, creativity, the dare to take risks, action-oriented, leadership, hard work, honesty, discipline, innovation, responsibility, cooperation, unyielding, realistic commitment, curiosity, communicative, and strong motivation for success. This integration is conducted in the learning process, through the role of the teacher as a designer, and the implementation is as outlined in the preparation of the syllabus and lesson plans.
Material and Methods
Preparation of termite nest sample Actinomycetes isolates growth
Sample of termite nest was collected in Sep- tember 2016 from Pananjung Pangandaran Na- ture Reserve, West Java, Indonesia. Termite nest was obtained by cruise method. The type of col- lected nest termite was carton nest. Termite nest samples (200 g per sample) were placed in poly- ethylene bags and immediately transported to Mi- crobiology Laboratorium of Research Center for Biomaterials- LIPI. Termites sample (worker and soldier) inhabiting the nest were collected and preserved in 70% alcohol tube. The termites were identified based on key identification. Photo- graphs were taken with digital microscope with 40 – 80 × magnification. Nest samples were ground into fine particles and air-dried at an am- bient temperature for 7 days before the isolation of Actinomycetes.
Figure 1. Actinomycetes isolates growth in HV agar media
Results and Discussion
Isolation of Actinomycetes from termite’s nest sample
In this research, we using HV agar media to isolate Actinomycetes from the termite’s nest sample. Its classify into nutrient-poor media therefore not many other microorganisms are able to live and utilize nutrients available in the media other than Actinomycetes bacterial group.
In the process of Actinomycetes.
JK6EM | Journal of K6, Education, and Management 280 Volume 2 | Number 3 | September | 2019
(a) (b)
Figure 2. Antibacterial activity of isolate Pn-TN2 against bacterial pathogen 1. E. coli, 2. S. aureus, 3. B. sub- tilis, 4. S. marcescens (a) and negative control of bacterial pathogen growth in ISP 2 media (b)
Table 1. Inhibition values and level of inhibition produced by isolated Actinomycetes against bacterial pathogen
Code isolate
Inhibition values (mm) and level of inhibition B.
subtili s
Level of inhibition
E. coli Level of inhibition
S.
marcescen s
Level of inhibition
P.
aeroginos a
Level of inhibition
Pn-TN1 0 - 0 - 0 - 0 -
Pn-TN2 32 +++ 30 +++ 0 - 41 +++
Pn-TN3 21 ++ 3 + 0 - 45 +++
Pn-TN4 0 - 0 - 0 - 0 -
Pn-TN5 10 + 0 - 0 - 3 +
Pn-TN6 7 + 0 - 0 - 4 +
Pn-TN7 4 + 4 + 0 - 0 -
Pn-TN8 9 + 7 + 0 - 6 +
Pn-TN9 0 - 0 - 0 - 5 +
Pn-TN10 0 - 0 - 0 - 0 -
Table 2. Growth of mycelia and inhibition rate produced by isolated Actinomycetes against fungal pathogen Code
Isolate
Growth of Mycelia (mm) Inhibition Rate (%)
F. palustris T. viridae F. oxysporum F.
palustris
T. viridae F.
oxysporum
a b a b a b
Pn-TN1 16.0 25.0 26.0 30.0 13.5 29.5 36.00 ±
9.3
13.33 ± 8.2 54.24 ± 3.5
Pn-TN2 10.0 26.0 10.5 28.0 6.0 30.0 61.54 ±
4.2
62.50 ± 4.4 80.00 ± 4.7
NPRA Krishanti, D Zulfiana, B Wikantyoso et al., 2018 / Antimicrobial Production by an Actinomycetes Isolate
ngal test. In the secondary bio-assay, we found contain in soil can be detected and avoided by termites with several disea-
se resistance mechanism. A trend in termite evo- lution of nests was observed progressively reduc- ing contact with soil. The high incidence of ento- mopathogens in soil were likely involved in the evolution of the more derived nesting habits in Nasutitermes. Environmental factors such as moisture, temperature, and light may be attrib- uted to characteristic of termite nesting then may lead to influence the composition of microbial communities in termite nest [41].
Conclusion and Recommendation
The strain S. prasinopilosus Pn-TN2 which isolated from termite nest sample, showed a broad range of antimicrobial activity against bac- terial and fungal test. Result showed that this iso- late possessed inhibition rate antifungal activity by 80% against F. oxysporum, 61% against F.
palustris, and 62% against T. viridae. Advised, this isolate also possessed high antibacterial ac- tivity against B. subtilis, P. aeroginosa, and E.
coli. Sequence analysis of 16S rRNA gene indi- cated that isolate Pn-TN2 belongs to genus Strep- tomyces. It shares 99% similarity with S.
prasinopilosus. We conclude that termite nest represent source of cultivable Actinomycetes that contribute diverse bio
active compounds for further biotechnology ap- plication in the medical, pharmaceutical, and agricultural fields.
Acknowledgment
This work was financially supported by Re- search Center for Biomaterials through “DIPA 2017”. Therefore, we are grateful for this funding and support of this research.
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